onium salts of taxo-diterpenoid-Cn,2-O-aza-arenes form water soluble self-assembling nanostructures above a critical aggregation concentration. The onium salt of aza-arene includes a delocalized charge which renders derivatized taxo-diterpenoids amphoteric, enhances their solubility in water, and promotes self-assembly of water soluble nanostructures, e.g., helical fibular structures and spherical micellular structures. These same onium salts of taxo-diterpenoid-Cn,2-O-aza-arenes are employed as water soluble prodrugs. For example, taxol-2'-methylpyridinium tosylate (MPT) is characterized by an elevated aqueous solubility, rapid activation by serum protein, good stability in most other aqueous solutions, formation of a protein:taxol intermediate and good retention within the circulatory system. The toxicity of the activated form is comparable or greater than underivatized taxol. Furthermore, taxol-2'-MPT can be synthesized by a simple one step reaction between taxol and 2-fluoro-1-MPT. The invention is applicable to taxol and taxol memetics having hydroxyls that are reactive with onium salts of 2-halogenated-aza-arenes. For example, taxol, C-2 substituted taxol and Taxotere each have reactive hydroxyls at the 2' and 7 positions. The invention is also applicable to a wide array of 2-halogenated-aza-arenes.
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6. A water soluble self-assembled micellular nanostructure having a spherical shape with a diameter of 20-120 Å and incorporating an onium salt of a taxo-diterpenoid-2-O-aza-arene.
4. A composition comprising:
an aqueous solvent and an onium salt of a taxo-diterpenoid-2-O-aza-arene dissolved in said aqueous solvent and capable of forming a water soluble self-assembled helical fibrous nanostructure above a critical aggregation concentration.
8. A composition comprising:
an aqueous solvent and an onium salt of a taxo-diterpenoid-2-O-aza-arene dissolved in said aqueous solvent and capable of forming a water soluble self-assembled micellular nanostructure above a critical aggregation concentration upon sonication.
10. A process for making a self-assembled micellular nanostructure from an aqueous solution including an onium salt of a taxo-diterpenoid-2-O-aza-arene, said process employing a step of sonicating the aqueous solution at a concentration in excess of a critical aggregation cencentration for the onium salt of the taxo-diterpenoid-2-O-aza-arene.
1. A water soluble self-assembled helical fibrous nanostructure having a diameter of 60-120 Å and a helical twist of 5-10 molecules per turn of an onium salt of a taxo-diterpenoid-2-O-aza-arene produced according to the following method:
step A: derivatizing a taxo-diterpenoid having a free hydroxyl with an onium salt of a 2-halogenated aza-arene for producing the taxo-diterpenoid-2-O-aza-arene; step B: forming the water soluble self-assembled helical fibrous nanostructure of the onium salt of the taxo-diterpenoid-2-O-aza-arene by solubilizing the taxo-diteroenoid-2-O-aza-arene of said step A within an aqueous solution and at concentration suitable for the formation of said water soluble self-assembled helical fibrous nanostructure.
22. A self-assembled helical fibrous nanostructure having a spherical shape with a diameter of 20-120 Å and incorporating an onium salt of a taxo-diterpenoid-Cn,2-O-aza-arene having an aqueous solubility in excees of 10-3 moles per liter, said onium salt of said taxo-diterpenoid-Cn,2-O-aza-arene represented by the following formula: ##STR49##
wherein: Cn is selected from the group consisting of C7 and C2' ; Rx is selected from the group consisting of Ph and tBuO; R10 is selected from the group consisting of OAc and OH; Ry is a C-2 substituent selected from the group consisting of benzyl and the following structures: ##STR50## R2' and R7 are each selected from the group consisting of OH and an onium salt of a 2-O-aza-arene, with the proviso that at least 2-O-aza-arene, said onium salt of the 2-O-aza-arene being selected from the group consisting of onium salt I and onium salt II represented by the following formulas: ##STR51## wherein: Z1 and Z2 are each selected from the group consisting of C and N; Z3 is selected from the group consisting of S and O; R1 is selected from the group consisting of C1 -C6 alkyl, allyl, arenxyl, propargyl, and fused aryl; R2 and R6 are each selected from the group consisting of H, C1 -C6 alkyl, allyl, arenxyl, propargyl, and fused aryl; if Z1 is C, then R3 is selected from the group consisting of H, C1 -C6 alkyl, allyl, arenxyl, propargyl, C1 -C6 O-alkyl, OH, halogen, and fused aryl; if Z1 is N, then R3 is absent; R4 and R8 are each selected from the group consisting of H, C1 -C6 alkyl, allyl, arenxyl, propargyl, C1 -C6 O-alkyl, OH, halogen, and fused aryl; and if Z2 is C, then R5 is selected from the group consisting of H, C1 -C6 alkyl, allyl, arenxyl, propargyl, C1 -C6 O-alkyl, OH, halogen, and fused aryl; if Z2 is N, then R5 is absent; and S- is a counter ion. 11. A self-assembled helical fibrous nanostructure having a diameter of 60-120 Å and a helical twist of 5-10 molecules per turn of an onium salt of a taxo-diterpenoid-Cn,2-O-aza-arene having an aqueous solubility in excees of 10-3 moles per liter, said onium salt of said taxo-diterpenoid-Cn,2-O-aza-arene represented by the following formula: ##STR46##
wherein: Cn is selected from the group consisting of C7 and C2' ; Rx is selected from the group consisting of Ph and tBuO; R10 is selected from the group consisting of OAc and OH; and Ry is C-2 substituent selected from the group consisting of benzyl and the following structures: ##STR47## R2' and R7 are each selected from the group consisting of OH and an onium salt of a 2-O-aza-arene, with the proviso that at least one of R2' and R7 is said onium salt of the 2-O-aza-arene, said onium salt of the 2-O-ara-arene, consisting of onium salt I and onium salt II represented by the following formulas: ##STR48## wherein: Z1 and Z2 are each selected from the group consisting of C and N; Z3 is selected from the group consisting of S and O; R1 is selected from the group consisting of C1 -C6 alkyl, allyl, arenxyl, propargyl, and fused aryl; R2 and R6 are each selected from the group consisting of H, C1 -C6 alkyl, allyl, arenxyl, propargyl, and fused aryl; if Z1 is C, then R3 is selected from the group consisting of H, C1 -C6 alkyl, allyl, arenxyl, propargyl, C1 -C6 O-alkyl, OH, halogen, and fused aryl; if Z1 is N, then R3 is absent; R4 and R8 are each selected from the group consisting of H, C1 -C6 alkyl, allyl, arenxyl, propargyl, C1 -C6 O-alkyl, OH, halogen, and fused aryl; and if Z2 is C, then R5 is selected from the group consisting of H, C1 -C6 alkyl, allyl, arenxyl, propargyl, C1 -C6 O-alkyl, OH, halogen, and fused aryl; if Z2 is N, then R5 is absent; and S- is a counter ion. 2. A water soluble self-assembled helical fibrous nanostructure as described in
3. A water soluble self-assembled helical fibrous nanostructure as described in
5. A composition as described in
said water soluble self-assembled helical fibrous nanostructures having a diameter of 60-120 Å and a helical twist of 5-10 molecules per turn.
7. A water soluble self-assembled micellular nanostructure as described in
9. A composition as described in
said water soluble self-assembled micellular nanostructure having a spherical shape with a diameter of 20-120 Å.
12. A self-assembled helical fibrous nanostructure as described in
S- is selected from the group of counter ions consisting of Cl-, Br-, I-, BF4-, ClO4-, ArSO3-, and C1 -C6 alkylSO3-.
13. A self-assembled helical fibrous nanostructure as described in
R2' is said onium salt and R7 is OH.
14. A self-assembled helical fibrous nanostructure as described in
Rx is Ph; R10 is AcO; and said onium salt is onium salt I wherein: Z1 and Z2 are both C; R1 is C1 -C6 alkyl; R2, R3, R4, and R4 are each H; and S- is selected from the group of counter ions consisting of Cl-, Br-, I-, BF4-, ClO4-, ArSO3-, and C1 -C6 alkylSO3-. 15. A self-assembled helical fibrous nanostructure as described in
R1 is methyl; and S- is ArSO3-.
16. A self-assembled helical fibrous nanostructure as described in
R2' and R7 are both said onium salt.
17. A self-assembled helical fibrous nanostructure as described in
Rx is Ph; R10 is AcO; and said onium salt is onium salt I wherein: Z1 and Z2 are both C; R1 is C1 -C6 alkyl; R2, R3, R4, and R4 are each H; and S- is selected from the group of counter ions consisting of Cl-, Br-, I-, BF4-, ClO4-, ArSO3-, and C1 -C6 alkylSO3-. 18. A self-assembled helical fibrous nanostructure as described in
R1 is methyl; and S- is ArSO3-.
19. A self-assembled helical fibrous nanostructure as described in
R7 is said onium salt and R2' is OH.
20. A self-assembled helical fibrous nanostructure as described in
Rx is Ph; R10 is AcO; and said onium salt is onium salt I wherein: Z1 and Z2 are both C; R1 is C1 -C6 alkyl; R2, R3, R4, and R4 are each H; and S- is selected from the group of counter ions consisting of Cl-, Br-, I-, BF4-, ClO4-, ArSO3-, and C1 -C6 alkylSO3-. 21. A self-assembled helical fibrous nanostructure as described in
R1 is methyl; and S- is ArSO3-.
23. A self-assembled helical fibrous nanostructure as described in
S- is selected from the group of counter ions consisting of Cl-, Br-, I-, BF4-, ClO4-, ArSO3-, and C1 -C6 alkylSO3-.
24. A self-assembled helical fibrous nanostructure as described in
R2' is said onium salt and R7 is OH.
25. A self-assembled helical fibrous nanostructure as described in
Rx is Ph; R10 is AcO; and said onium salt is onium salt I wherein: Z1 and Z2 are both C; R1 is C1 -C6 alkyl; R2, R3, R4, and R4 are each H; and S- is selected from the group of counter ions consisting of Cl-, Br-, I-, BF4-, ClO4-, ArSO3-, and C1 -C6 alkylSO3-. 26. A self-assembled helical fibrous nanostructure as described in
R1 is methyl; and S- is ArSO3-.
27. A self-assembled helical fibrous nanostructure as described in
R2' and R7 are both said onium salt.
28. A self-assembled helical fibrous nanostructure as described in
Rx is Ph; R10 is AcO; and said onium salt is onium salt I wherein: Z1 and Z2 are both C; R1 is C1 -C6 alkyl; R2, R3, R4, and R4 are each H; and S- is selected from the group of counter ions consisting of Cl-, Br-, I-, BF4-, ClO4-, ArSO3-, and C1 -C6 alkylSO3-. 29. A self-assembled helical fibrous nanostructure as described in
R1 is methyl; and S- is ArSO3-.
30. A self-assembled helical fibrous nanostructure as described in
R7 is said onium salt and R2' is OH.
31. A self-assembled helical fibrous nanostructure as described in
Rx is Ph; R10 is AcO; and said onium salt is onium salt I wherein: Z1 and Z2 are both C; R1 is C1 -C6 alkyl; R2, R3, R4, and R4 are each H; and S- is selected from the group of counter ions consisting of Cl-, Br-, I-, BF4-, ClO4-, ArSO3-, and C1 -C6 alkylSO3-. 32. A self-assembled helical fibrous nanostructure as described in
R1 is methyl; and S- is ArSO3-.
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The invention disclosed herein was supported in part by Grant Number CA 46446-CH from the National Institutes of Health. The United States government may have certain rights to this invention.
The present application is a 371 of PCT patent application PCT/US95/00538, filed Jan. 11, 1995, which is a continuation of U.S. patent application Ser. No. 08/180,136, filed Jan. 11, 1994, now abandoned, the disclosures of which is incorporated herein by reference.
1. Technical Field
The invention relates to taxol prodrugs. More particularly, the invention relates to taxol and taxol memetics derivatized with onium salts of aza-arenes to form water soluble prodrugs which form stable self-assembled nanostructures in aqueous solvents having helical and micellular structures and which are converted to active form upon contact with serum protein.
2. Background
Taxol, an antineoplastic agent originally isolated from Taxus brevifolia, is approved for usage in the treatment of ovarian cancer and is expected to see usage in breast, lung, and skin cancers as well. However, since Taxol possesses an extremely low water solubility, i.e., less than 1.5×10-6 M, it has been necessary to formulate Taxol in a mixture of Cremaphor™, a polyoxyethylated castor oil, and ethanol in order to achieve a therapeutic concentration. This formulation can induce a variety of significant side effects including hypersensitivity reactions.
While premedication and slow administration of the drug can circumvent these problems in the clinic, the entire protocol is quite cumbersome and requires extensive close monitoring of patients. Although taxol's dramatic efficacy has driven clinical usage forward despite these problems, a water soluble form of taxol could completely obviate the need for this troublesome protocol.
One approach to bypassing these formulation difficulties, previously attempted by several groups including our own, is the introduction of solubilizing functionality that normal metabolic pathways could remove in vivo. Compounds of this type, termed prodrugs, consist, in the case of taxol, primarily of ester derivatives at the 2' and 7 positions. Currently none of these protaxols have given success in the clinic. In each case, the prodrug is rapidly cleared from circulation by the kidneys.
Taxol is only one of a class of taxo-diterpenoids having bioactivity. Another preferred taxo-diterpenoid having clinically significant activity is Taxoter™. Unfortunately, all known bioactive taxo-diterpenoids have a low aqueous solubility.
What is needed is a stable prodrug which is activated by contact with serum and which is retained in circulation for a clinically significant period after administration.
The invention is directed to water soluble self-assembling nanostructures of onium salts of taxo-diterpenoid-Cn, 2-O-aza-arenes and solutions thereof. The onium salt of aza-arene includes a delocalized charge which renders the molecule amphoteric and enhances its solubility in water. In a first embodiment, the invention herein teaches that water soluble onium salts of taxo-diterpenoid-Cn,2-O-aza-arenes self-assemble in aqueous media to form water soluble helical nanostructure. In a second embodiment, the invention herein teaches that, with the aid of sonication, water soluble onium salts of taxo-diterpenoid-Cn,2-O-aza-arenes self-assemble in aqueous media to form water soluble micellular nanostructure. The invention further teaches that these nanostructures are stable in aqueous media over a period of hours or days. It is also taught that, with the addition of serum protein, 2-O-aza-arene is displaced from the onium salt of taxo-diterpenoid-Cn,2-O-aza-arene and a soluble protein:taxo-diterpenoid intermediate is formed. Furthermore, it is taught herein that the protein:taxo-diterpenoid intermediate dissociates over time to provide a bioactive taxo-diterpenoid. Preferred taxo-diterpenoids include taxol, C-2 substituted taxol analogs, and Taxotere™. Taxo-diterpenoid-Cn,2-O-aza-arene may be produced in a one step synthesis by reacting onium salts of 2-halogenated aza-arenes with reactive hydroxyls on the taxo-diterpenoid. Reactive hydroxyls on taxol and Taxotere™ are located at C2 ' and C7. A preferred onium salt of 2-halogenated aza-arene is 2-fluoro-1-methylpyridinium tosylate. Other employable onium salts of 2-halogenated aza-arenes are disclosed by T. Mukaiyama, Angewandte Chemie 1979, 18(18), 707-808, incorporated herein by reference.
More particularly, the first embodiment of the invention is directed to water soluble self-assembled helical fibrous nanostructure having a diameter of 60-120 Å and a helical twist of 5-10 molecules per turn of an onium salt of a taxo-diterpenoid-2-O-aza-arene wherein the solubility of the onium salt of the taxo-diterpenoid-2-O-aza-arene exceeds 10-3 moles per liter and the 2-O-aza-arene is labile to displacement by serum protein. The first embodiment of the invention is also directed to compositions comprising an aqueous solvent in combination with dissolved taxo-diterpenoid-2-O-aza-arenes capable of forming such water soluble self-assembled helical fibrous nanostructures above a critical aggregation concentration.
A second embodiment of the invention is directed to water soluble self-assembled micellular nanostructures having a spherical shape with a diameter of 20-120 Å and incorporating onium salts of taxo-diterpenoid-2-O-aza-arenes wherein the solubility of the onium salt of the taxo-diterpenoid-2-O-aza-arene exceeds 10-3 moles per liter. The second embodiment of the invention is also directed to compositions comprising an aqueous solvent in combination with dissolved taxo-diterpenoid-2-O-aza-arenes capable of forming water soluble self-assembled micellular nanostructures above a critical aggregation concentration upon sonication. The invention is also directed to a process for making self-assembled micellular nanostructures from an aqueous solution including an onium salt of a taxo-diterpenoid-2-O-aza-arene. This process employs sonication at a concentration in excess of a critical aggcentration for the onium salt of the taxo-diterpenoid-2-O-aza-arene. In a preferred embodiment, the taxo-diterpenoid-2-O-aza-arenes are represented by the following formula: ##STR1##
wherein Rx is Ph or tBuO; R10 is OAc or OH; Ry is a C-2 substituent defined below; and R2 ' and R7 are each selected from the group consisting of OH and an onium salt of a 2-O-aza-arene, with the proviso that at least one of R2 ' and R7 is the onium salt of the 2-O-aza-arene. The onium salt of the 2-O-aza-arene can be represented by either of the following formulas for onium salt I or onium salt II: ##STR2##
wherein Z1 and Z2 are each either C or N; Z3 is S or O; R1 is selected from the group consisting of C1 -C6 alkyl, allyl, arenxyl, propargyl, and fused aryl; R2 and R6 are each selected from the group consisting of H, C1 -C6 alkyl, allyl, arenxyl, propargyl, and fused aryl; if Z1 is C, then R3 is selected from the group consisting of H, C1 -C6 alkyl, allyl, arenxyl, propargyl, C1 -C6 O-alkyl, OH, halogen, and fused aryl; if Z1 is N, then R3 is absent; R4 and R8 are each selected from the group consisting of H, C1 -C6 alkyl, allyl, arenxyl, propargyl, C1 -C6 O-alkyl, OH, halogen, and fused aryl; and if Z2 is C, then R5 is selected from the group consisting of H, C1 -C6 alkyl, allyl, arenxyl, propargyl, C1 -C6 O-alkyl, OH, halogen, and fused aryl; if Z2 is N, then R5 is absent; and S- is a counter ion.
Preferred C-2 Ry substitution include phenyl or the following: ##STR3##
FIG. 1 illustrates the kinetics of taxol release from taxol-2'-MPT (2) in various aqueous solutions at 25°C (horizontal line). Although stable in water and aqueous buffer solutions, methylene chloride extraction of plasma treated with compound 2 showed complete conversion of 2 into taxol (1) within 10 minutes (curve, 20% of total recovered).
FIG. 2A illustrates a transmission electron micrograph (TEM) of self-assembled helical fibrous nanostructures of taxol-2'-MPT, i.e., compound 2 in buffered solutions (2,1 mM in 100 mM PBS) above the critical aggregation concentration (CAC) of this compound using a negative phosphotungstate stain and a magnification of ×25000. The inset shows a portion of one of the fibrils further magnified to illustrate the helical nature of the structure.
FIG. 2B illustrates a transmission electron micrograph (TEM) of self-assembled spherical nanostructures of taxol-2'-MPT (compound 2) in unbuffered solutions (2,1 mM in H2 O) above the critical aggregation concentration (CAC) of this compound using a negative uranyl acetate stain and a magnification of ×45000.
FIG. 3 illustrates a tubulin polymerization-depolymerization measurements with negative control (triangles), positive taxol control (diamonds), and taxol-2'-MPT, i.e., compound 2 (dots). Calcium chloride promoted depolymerization is suppressed by taxol but not by taxol-2'-MPT.
FIG. 4 illustrates the relative cytotoxicities of taxol-2'-MPT (compound 2) and taxol against a variety of cell lines.
FIG. 5 illustrates the efficacy of taxol-2'-MPT in lung tumor xenograft nude mouse model: 5% dextrose (triangles), taxol (diamonds), and taxol-2'-MPT (dots).
The synthesis, physical properties, and pharmaceutical profiles of water soluble onium salts of taxo-diterpenoid-Cn,2-O-aza-arenes are described.
Taxol-2'-MPT (methylpyridinium tosylate), compound 2, was synthesized according to the method of T. Mukaiyama, Angewandte Chemie 1979, 18(18), 707-808, incorporated herein by reference. Taxol (10 mg, 0.012 mM), from NaPro Biochemicals, Boulder, Colo., USA, was dried by azeotropic distillation with toluene (2×1.0 mL) and then dissolved in methylene chloride (0.4 mL) and treated sequentially under an atmosphere of dry argon, with freshly distilled triethylamine (5 microl, 0.04 mM, 3 equivalents) and 2-fluoro-1-methylpyridinium tosylate (5 mg, 0.018 mM, 1.5 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 30 minutes. The clear colorless solution rapidly turned to a clear pale yellow. The course of the reaction was monitored through thin layer chromatography (TLC) (E. Merck RP-18 silica, 65 tetrahydrofuran: 35 water, UV/phosphomolybdic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC (Rf 0.8). Purification via reverse phase high pressure liquid chromatography (HPLC) (C18 column, 1 mM NH4 OAc pH 6.5 buffer/methanol gradient, 1.5 mL/min. UV) to give, after removal of solvent in vaccuo, pure taxol-2'-MPT (2) (12 mg, 93% yield) as a white amorphous solid. All spectroscopic data (1 H NMR and HRMS) were in accord with the structure assigned to 2. 1 H NMR (CDCl3, 125 MHz) δ: 1.055 (s, 3 H, C17-H), 1.083 (s, 3 H, C19-H), 1.724 (s, 3 H, C19-H), 1.858 (m, 1 H, C6-_H), 1.913 (s, 3 H, CH3 -Ph), 2.193 (s, 3 H, C10-OC(O)CH3), 2.514 (m, 1 H, C6-aH), 3.663 (d, 1 H, J=7.0 Hz, C3-H), 4.110 (d, 1 H, J=8.5, C20-_H, A of AB), 4.133 (s, 3 H, N-CH3), 4.230 (d, 1 H, J=8.5, C20-aH, B of AB), 4.315 (dd, 1 H, J=8.7, 10.7, C7-H), 4.901 (dd, 1 H, J=1.0, 7.7, C5-H), 5.501 (d, 1 H, C2-H, J=7.0), 5.702 (bt, 1 H, C2'-H, J=8.0), 5.951 (dd, 1 H, C13-H, J=1.0, 8.0), 6.120 (bd, 1 H, C3'-H, J=10.0), 6.181 (s, 1 H, C10-H) 7.702 (t, 1 H, N-H, J=7.5), 7.33-7.45 (m, 8 H, Ar-H), 7.56-7.62 (m, 4 H, Ar-H), 7.56-7.62 (m, 4 H, Ar-H), 7.68-7.75 (m, 4 H, Ar-H), 8.00-8.05 (m, 1 H Ar-H), 8.23-8.28 (m, 1 H, Pyr-H), 8.41 (m, 1 H, Pyr-H). IR (neat, KCl plate) cm-1 : 3640-3120 (bm), 3030-2870 (bm), 2320 (m), 1720 (s), 1630 (m), 1560 (m), 1500 (m), 1360 (s), 1160 (m), 1070 (m), 700 (m). UV/Vis (CHCl3) nm: 254, 280. FAB HRMS: calc for C53 H57 O14 N2 : 945.3810; found: 945.3810.
The molecular structures of taxol, compound 1, and of taxol-2'-MPT, compound 2, are illustated in Scheme 1A. The synthesis of taxol-2'-MPT is illustrated in Scheme 1B. ##STR4##
The synthesis of taxol-7-MPT differed only slightly from the synthesis of Taxol-2'-MPT. Taxol (10 mg, 0.012 mM), from NaPro Biochemicals, Boulder, Colo., USA, was dissolved in methylene chloride (2.0 mL) and treated sequentially with triethylamine (67 microL, 0.48 mM, 40 equivalents) and 2-fluoro-1-methylpyridinium tosylate (34 mg, 0.12 mM, 10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 5 minutes. Purification via reverse phase high pressure liquid chromatography (HPLC) gave pure taxol-2'-MPT (2) (12 mg, 93% yield) as a white amorphous solid. The Rf of taxol-7-MPT is about 0.3 minutes less than the Rf of taxol-2'-MPT. The yield was 11 mg or 85%. Spectroscopic data (1 H NMR and HRMS) were as expected.
The synthesis of taxol-bis-2',7-MPT differed from the synthesis of Taxol-7-MPT only with respect to reaction time. Taxol (10 mg, 0.012 mM), from NaPro Biochemicals, Boulder, Colo., USA, was dissolved in methylene chloride (2.0 mL) and treated sequentially with triethylamine (67 microL, 0.48 mM, 40 equivalents) and 2-fluoro-1-methylpyridinium tosylate (34 mg, 0.12 mM, 10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 18 hours. Purification via reverse phase high pressure liquid chromatography (HPLC) gave pure taxol-2'-MPT (2) (12 mg, 93% yield) as a white amorphous solid. The Rf of taxol-bis-2',7-MPT is about 0.3 minutes less than the Rf of taxol-2'-MPT. The yield was 13 mg or 85%. Spectroscopic data (1 H NMR and HRMS) were as expected.
Alternative synthetic schemes based upon the method of T. Mukaiyama (Angewandte Chemie 1979, 18(18), 707-808) using a variety of onium salts of 2-halogenated aza-arenes for derivatizing either the 2' or the 7 positions of taxol are illustrated in the following scheme: ##STR5##
Due to a difference in retention time using our standard HPLC conditions (see FIG. 1) and differing ultraviolet absorption maxima (l280 /l254 =1.6 for 2 and 0.3 for 1), the stability of 2 was easily assayed by HPLC (FIG. 1). In all of the ensuing studies, the only degradation products detected were taxol and the pyridinone that results from hydrolysis of pyridinium salts (FIG. 1c.). Taxol-2'-MPT appears completely stable in the solid state in a temperature range of -80° C. to 25°C regardless of the presence of an inert atmosphere. In water, 5% dextrose, and 1.5% saline 2 is stable for several days but begins to exhibit slow degradation after 4 days. In phosphate buffered saline (PBS) or ammonium acetate-phosphate buffer systems of pH 6.0 to 7.3, 2 is stable at 25°C for over 21 days. Taxol-2'-MPT (2) is, however, unstable in 5% HCl (pH 1.1) and brine. Most significantly, 2 breaks down rapidly when incubated at 37°C with human plasma. This result suggests the presence of factors within plasma that initiate the degradation of 2 to taxol. Since taxol has been shown to bind to albumin to the extent of ca. 85% in sera, it is suspected that basic lysine residues on this protein may initiate breakdown.
The kinetics of taxol release from taxol-2'-MPT (2) in various aqueous solutions at 25°C is shown in FIG. 1. In sterile water, pH 6.2 phosphate buffered saline, or 5% dextrose, no taxol release is observed over a period of 11 minutes, as shown by the horizontal line. Although stable in water and aqueous buffer solutions, methylene chloride extraction of plasma treated with compound 2 showed complete conversion of 2 into taxol (1) within 10 minutes, as shown by the curved line. Under these conditions 20% of total is recovered. More particularly, Taxol-2'-MPT (2) was dissolved in the aqueous system with the aid of sonication for five minutes. Aliquots were then removed at the times shown and partitioned into methylene chloride to quench the reaction. Samples were then analyzed using a Waters Maxima HPLC instrument equipped with an autoinjector (3.9×300 mm C18 column equipped with a precolumn. The flow rate was 1.5 mL/minute. The eluent gradient A-B extended over 30 minutes. "A" was 80% 80 mM ammonium acetate, pH 6∅ "B" was 100% methanol. An ultraviolet diode array detector was employed. The ratio of compound 2 (Rf 16.2 min.) to taxol (compound 1, Rf 16.8 min.) remaining was determined from the relative areas of the peaks after normalization with previously determined calibration curves.
The solubility and partition coefficient data for 2 and taxol were determined using an HPLC method.
TBL Taxol 2'-MPT-taxol Solubility in Water <1.5 × 10-6 1.7 × 10-3 Partition Coefficient >10000 50 [octanol/[water]Solubility was found by forming a solution in water with the aid of sonication for five minutes, centrifugation of the samples, and injection of the supernatant. The values reported were normalized using calibration curves constructed for both compounds by preparing known concentrations in the range 1×10-6 to 1×10-3 M in methylene chloride and subjecting to HPLC analysis under the same conditions. One should note that the solubility of taxol is at the detection limit of the instrument and thus represents an upper limit. The solubility of 2 was found at a concentration at which the solution was clear (see below) and thus represents a lower limit. Compound 2 exhibited similar solubilities for various buffer systems in the pH range 6.2 to 7.4. Partition coefficients were determined by dissolving the compound in the organic phase, shaking the resulting solution with water for ten minutes, and analyzing each phase by HPLC as above. No degradation of 2 was noted during these studies.
These data clearly show that 2 is significantly more soluble in water than the parent taxol. The solubility demonstrated in a range of aqueous systems is higher than the clinically relevant dosages (3 to 30 mM).
While the water solutions of 2 were optically clear at all concentrations examined, buffered solutions of concentrations greater than 1×10-3 M exhibited a haze to the naked eye and diffuse scattering of monochromatic light. Ultraviolet spectroscopic absorption measurements at 340 nm (FIG. 3) showed an exponential increase in optical density (OD) above a critical concentration of 4×10-4 M, a result characteristic of macromolecular structure in solution. Transmission electron microscopy (TEM) confirmed the presence of supramolecular structures in these solutions. Uniform aggregates of fibrillar structure (FIG. 2a) with helical conformations were observed. These structures exhibited varying (up to 800 Å) lengths but consistent diameter of ca. 80 Å with a helical twist of about 7. Additionally, freshly sonicated solutions of 2 showed the presence of spherical structures (FIG. 2b) with diameters of about 50 Å. It is likely that the long term stability of these solutions is due, at least in part, to stabilization provided by this structured environment.
Microtubule polymerization-depolymerization measurements (FIG. 3) with taxol-2'-MPT (2) were very similar to GTP-saline controls and drastically different from taxol. Compound 2 does not appear to bind to tubulin in the manner of taxol. In the buffered aqueous environment of this assay, 2 is not converted to taxol and thus does not affect the tubulin-microtubule equilibria. Taxol, recovered from human plasma treated with 2, exhibited the expected microtubule stabilization, indicating that 2 does act as a prodrug for taxol.
Tubulin polymerization-depolymerization measurements are illustrate in FIG. 3. Negative controls are shown with triangles; positive taxol controls are shown with diamonds; and taxol-2'-MPT, i.e., compound 2 is shown with dots. The measurements indicate that calcium chloride promoted depolymerization is suppressed by taxol but not by taxol-2'-MPT.
More particularly, measurement were performed in 96 well plates at 37°C following the protocol of R. Merlock and W. Wrasidlo (Analytical Biochemistry 1993, in press). Calcium chloride addition is indicated by the arrow. In each case, 1.0 mM GTP was used to promote the initial polymerization of tubulin. Negative control employed tubulin (1.0 mg/mL) alone, CaCl2 (0.25 mM) added after 20 minutes. Positive taxol control employed tubulin (1.0 mg/mL) with taxol (10-6 M) and CaCl2 (0.25 mM) added after 20 minutes. The experimental taxol-2'-MPT employed tubulin (1.0 mg/mL) with taxol-2'-MPT (10-6) and CaCl2 (0.25 mM) added after 20 minutes. Turbidity was measured as optical density at 340 nm using a microplate reader (Molecular Devices Thermomax).
Compound 2 was tested for its cytotoxicity against a cell line panel including leukemia, ovarian, lung, and breast carcinoma cells (FIG. 4). The differential cytotoxicity profiles for 2 and taxol were similar, although some differences were noted. Both compounds exhibited IC50 values ranging from 10-5 to 10-12 M with means close to one nanomolar. Normal cells had cytotoxicity levels three to four orders of magnitude below mean values. Extremely high cytotoxicity levels were recorded for human leukemia, metastatic melanoma and cervical carcinoma. As expected for a prodrug of taxol in the cellular environment, 2 shows the same remarkable tumor cell selectivity and cell line specificity as taxol.
The relative cytotoxicities of taxol-2'-MPT (compound 2) and taxol against a variety of cell lines are illustrated in FIG. 4. More particularly, cells were plated on 96 well plates with the following controls: no cells and toxic control (1×10-3 M SDS). The drug was added to the first set of wells and diluted via standard dilution method from the stock. Plates were incubated at 37°C, 5% CO2 in sterile air in an humidified incubator for 72 hours. An aliquot of 50 μL of a solution of 2,3-bis(methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-1H-tetraz olium hydroxide (XTT), 1 mg mL-1, in phosphate buffered saline (PBS, 100 mM) was added to the wells. In the presence of viable cells, this colorless clear media is enzymatically altered to give a pink coloration. The plates were read at 450 nm using a plate reader. Percentage cytotoxicity was calculated using the formula: %C=1-(OD toxin)(OD growth control)-1 (100).
The encouraging in vitro data obtained with taxol-2'-MPT (2) prompted us to study its in vivo action using nude mice inflicted with human lung carcinoma xenografts (FIG. 1). The samples of 2 used for this study were formulated in sterile PBS without Cremaphor™, indicating the suitability of this compound for simple bolus administration. Preliminary data shows that the control of tumor growth exhibited by 2 is at least comparable to that of taxol and significantly (0.001 p-value, multiple linear regression model) different from controls. These results provide a reasonable indication that 2 is converted rapidly to taxol in vivo and should thus exhibit pharmacology similar to taxol. Indeed, in metabolic study using tritiated 2, only 5% of the compound was excreted through the kidneys, a result that is completely in accord with the behavior of taxol.
The efficacy of taxol-2'-MPT in lung tumor xenograft nude mouse model is illustrated in FIG. 5. The tumor model was generated from an ATCC A549 non-small cell lung adenocarcinoma cell line that was maintained under the standard cell proliferation conditions (37°C, 5% carbon dioxide in sterile air). Hemocytometer counted cells suspended in Hanks medium (Gibco, Grand Island, N.Y.) were implanted S.C. (106 cells in 0.4 mL per tumor volume determined using the equation (length)(width)2 /2. The test compounds (1.0 microM) were injected I.P. on day 1,3, and 7 using the following media: control. 5% dextrose in water (D5W), triangles; taxol, suspended in Cremaphor/D5W (5/95, 18.0 mg/kg of animal weight), diamonds; and taxol-2'-MPT, dissolved in D5W (23.9 mg/kg of animal weight), dots. The procedures used for the maintenance of tumors and the experimental details were according to protocols set forth by the Developmental Therapeutics Program, National Cancer Institute, viz., National Cancer Institute Cancer Chemnotherapy Reports, 3 (1972).
The mechanism of acid catalyzed taxol (1) release from taxol-2'-MPT is illustrated in Scheme 2. ##STR6##
However, the release of taxol from taxol-2'-MPT can also be catalyzed by serum protein and by proteins having nucleophilic groups. When contacted with serum protein, taxol-2'-MPT is observed to displace its MPT group and form a protein:taxol intermediate. Dialysis of the protein:taxol intermediate indicates a dissociation period of hours or days. Displacement of the MPT group by serum proteins seems to be specific for such serum proteins. Tested non-serum proteins seemed to lack this activity. In particular, immunoglobulins and serum albumen seem to be particularly effective displacing the MPT group and forming protein:taxol intermediates. The precise nature of the bonding between the protein and taxol has not been characterized. Scheme 2 illustrates alternative pathways for MPT release.
Taxol-2'-MPT has proven to be a remarkably stable compound in most aqueous media. It is probable that this stability is conferred upon 2 by the facile formation of supramolecular aggregates, a process that is probably driven by the amphiphillic nature of the compound. The stability, water solubility, and lack of cytotoxicity of taxol-2'-MPT makes this class of compound an ideal a prodrug for taxol and memetics of taxol. While essentially completely stable in aqueous media at physiological pH and ion strength, the compound rapidly discharges taxol in sera. This profile is ideal for a clinically useful prodrug to taxol. It is possible that these properties should allow the formulation of taxol-2'-MPT (2) without the use of Cremaphor™ or ethanol. ##STR7##
PAC Preparation of 7-TES Deacetylbaccatin III (4)##STR8##
7-TES deacetylbaccatin III (4). To a solution of 10-deacetylbaccatin III (3, 3.0 g, 5.51 mmol, Indena Corpation, Italy) in pyridine (250 mL) was added chlorotriethylsilane (18.5 mL, 110 mmol) dropwise. The resulting solution was stirred at 25°C for 17 hours. After dilution with diethylether (750 mL), the solution was washed with aqueous CuSO4 (3×200 mL) and brine (200 mL). The organic layer was dried (MgSO4), concentrated, and purified by flash chromatography (silica, 35→50% ethylacetate in petroleum ether) to give alcohol 4 (3.39 g, 91%) as a white solid.
Physical Data for 7-TES deacetylbaccatin III (4). Rf =0.32 (silica, 50% ethylacetate in hexanes); IR (thin film) νmax 3464, 2954, 2282, 1710, 1453, 1362, 1271, 1242, 1105, 994 cm-1 ; 1 H NMR (500 MHz, CDCl3)δ 8.06 (dd, J=8.0, 0.9 Hz, 2 H, Bz), 7.57 (t, J=7.9 Hz, 1 H, Bz), 7.44 (t, J=7.9 Hz, 2 H, Bz), 5.56 (d, J=7.0 Hz, 1 H, 2-H), 5.14 (d, J=1.9 Hz, 1 H, 10-H), 4.92 (d, J=9.5 Hz, 1 H, 5-H), 4.84-4.78 (m, 1 H, 13-H), 4.37 (dd, J=10.6, 7.0 Hz, 1 H, 7-H), 4.27 (d, J=8.5 Hz, 1 H, 20-H), 4.25 (d, J=1.9 Hz, 1 H, 10-OH), 4.12 (d, J=8.5 Hz, 1 H, 20-H), 3.91 (d, J=7.0 Hz, 1 H, 3-H), 2.48-2.40 (m, 1 H, 6-H), 2.25 (s, 3 H, Me), 2.25-2.17 (m, 2 H, 14-CH2), 2.04 (s, 3 H, Me), 1.90-1.82 (m, 1 H, 6-H), 1.70 (s, 3 H, Me), 1.03 (s, 6 H, Me, Me), 0.90 (t, J=8 Hz, 9 H, Si(CH2 CH3)3), 0.58-0.42 (band, 6 H, Si(CH2 CH3)3); 13 C NMR (125 MHz, CDCl3) δ 210.3, 170.8, 167.0, 141.8, 135.1, 133.6, 130.1, 129.4, 128.6, 84.2, 80.7, 78.8, 76.5, 74.8, 74.6, 72.9, 67.9, 57.9, 47.0, 42.7, 38.6, 37.2, 26.8, 22.6, 19.5, 15.2, 9.9, 6.7, 5.1; FAB HRMS (NBA/CsI) m/e 791.2251, M+Cs30 calcd for C35 H50 O10 Si 791.2228.
##STR9##
Enone 5. To a solution of 7-TES deacetylbaccatin III (4, 1.5 g, 2.28 mmol) and 4-methylmorpholine N-oxide (NMO, 240 mg, 2.05 mmol) in CH2 Cl2 (5 mL) was added 4 Å molecular sieves (200 mg) and the suspension was stirred at 25°C for 10 minutesutes. A catalytic amount of tetrapropylammonium perruthenate from Aldrich Chemical Company Inc. (TPAP, 40 mg, 0.11 mmol) was added by portions and the reaction mixture was stirred at 25°C for 0.5 hours. Small amounts of 4-methylmorpholine N-oxide and TPAP were added alternatively at 0.5 hour intervals until the starting material was consumed to the extent of ca. 95% by TLC. The reaction mixture was filtered through silica gel, eluted with CH2 Cl2 (100 mL), and concentrated to give enone 5 (1.44 g, 96%) as a white solid.
Physical Data for Enone 5. Rf =0.5 (silica, 50% ethylacetate in hexanes); IR (thin film) νmax 3446, 2957, 2882, 1726, 1672, 1456, 1367, 1243, 1106 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.05 (dd, J=8.0, 1.0 Hz, 2 H, Bz), 7.61 (t, J=7.5 Hz, 1 H, Bz), 7.45 (t, J=7.5 Hz, 2 H, Bz), 5.63 (d, J=7.5 Hz, 1 H, 2-H), 5.30 (d, J=2.0 Hz, 1 H, 10-H), 4.90 (d, J=8.0 Hz, 1 H, 5-H), 4.36 (dd, J=10.5, 7.0 Hz, 1 H, 7-H), 4.31 (d, J=8.5 Hz, 1 H, 20-H), 4.30 (d, J=2.0 Hz, 1 H, 10-OH), 4.11 (d, J=8.5 Hz, 1 H, 20-H), 3.93 (d, J=7.5 Hz, 1 H, 3-H), 2.92 (d, J=19.5 Hz, 1 H, 14-H), 2.62 (d, J=19.5 Hz, 1 H, 14-H), 2.50-2.42 (m, 1 H, 6-H), 2.17 (s, 3 H, Me), 2.08 (s, 3 H, Me), 1.90-1.82 (m, 1 H, 6-H), 1.77 (s, 1 H, 1-OH), 1.70 (s, 3 H, Me), 1.21 (s, 3 H, Me), 1.14 (s, 3 H, Me), 0.90 (t, J=8.0 Hz, 9 H, Si(CH2 CH3)3), 0.60-0.42 (band, 6 H, Si(CH2 CH3)3); 13 C NMR (125 MHz, CDCl3) δ 208.2, 198.1, 170.2, 166.8, 156.6, 139.1, 134.0, 130.0, 128.8, 128.8, 84.0, 80.4, 78.5, 76.2, 75.7, 72.9, 72.8, 58.8, 45.9, 43.4, 42.5, 37.2, 33.0, 21.7, 17.5, 13.6, 9.6, 6.7, 5.1; FAB HRMS (NBA/NaI) m/e 657.3070, M+Na+ calcd for C35 H48 O10 Si 657.3095.
##STR10##
Triol 6. To a solution of enone 5 (1.44 g, 2.19 mmol) in MeOH (300 mL) at 0°C was slowly added an aqueous solution of K2 CO3 (3.0 g in 32 mL of H2 O). The solution was stirred at 0°C for 2.5 hours. The reaction was then quenched with aqueous NH4 Cl (150 mL) and the resulting mixture was extracted with CH2 Cl2 (2×200 mL). The organic layer was dried (Na2 SO4), concentrated, and purified by flash chromatography (silica, 35→50% ethylacetate in petroleum ether) to give enone 5 (270 mg, 19%) and triol 6 (912 mg, 93% based on 81% conversion).
Physical Data for Triol 6. Rf =0.24 (silica, 50% ethylacetate in hexanes); IR (thin film) νmax 3414, 2957, 2881, 1727, 1664, 1370 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 5.23 (d, J=9.5 Hz, 1 H, 10-H), 4.89 (d, J=9.5 Hz, 1 H, 5-H), 4.63 (d, J=9.5 Hz, 1 H, 20-H), 4.56 (d, J=9.5 Hz, 1 H, 20-H), 4.32 (dd, J=11.0, 7.0 Hz, 1 H, 7-H), 4.28 (d, J=2.5 Hz, 1 H, 10-OH), 3.89 (dd, J=6.5, 4.0 Hz, 1 H, 2-H), 3.57 (d, J=6.5 Hz, 1 H, 3-H), 2.78 (d, J=19.5 Hz, 1 H, 14-H), 2.58 (d, 4.0 Hz, 1 H, 2-OH), 2.52 (d, J=19.5 Hz, 1 H, 14-H), 2.49-2.42 (m, 1 H, 6-H), 2.03 (s, 3 H, Me), 1.92-1.84 (m, 1 H, 6-H), 1.68 (s, 3 H, Me), 1.21 (s, 3 H, Me), 1.04 (s, 3 H, Me), 0.90 (t, J=8.0 Hz, 9 H, Si(CH2 CH3)3), 0.60-0.40 (band, 6 H, Si(CH2 CH3)3); 13 C NMR (125 MHz, CDCl3) δ 208.9, 198.5, 170.1, 156.7, 138.8, 83.8, 81.2, 77.6, 75.7, 72.8, 72.5, 58.8, 45.8, 43.1, 42.8, 37.3, 32.7, 21.6, 17.5, 13.6, 9.7, 6.7, 5.1; FAB HRMS (NBA/NaI) m/e 575.2648, M+Na+ calcd for C28 H44 O9 Si 575.2652.
##STR11##
Carbonate 7. A solution of triol 6 (60.0 mg, 0.109 mmol) in THF (2 mL) was treated with carbonyldiimidazole (110.0 mg, 0.678 mmol) and stirred at 40°C for 0.5 hour The reaction mixture was concentrated and redisolved in THF (5 mL). TLC analysis confirmed total consumption of starting material. 1N aqueous HCl (5 mL) was added and the resulting solution was allowed to stir for 15 minutes at 25°C diethylether (25 mL) was added, the organic layer was separated, washed with aqueous NaHCO3 (10 mL) and brine (10 mL), dried (MgSO4), and concentrated to give carbonate 7 (58 mg, 93%) as a white foam.
Physical Data for Carbonate 7. Rf =0.50 (silica, 35% ethylacetate in hexanes); IR (thin film) νmax 3438, 2957, 2882, 1820, 1731, 1685, 1370, 1236 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 5.27 (d, J=2.5 Hz, 1 H, 10-H), 4.89 (d, J=9.0 Hz, 1 H, 5-H), 4.60 (d, J=9.0 Hz, 1 H, 20-H), 4.45 (d, J=9.0 Hz, 1 H, 20-H), 4.43 (d, J=6.0 Hz, 1 H, 2-H), 4.33 (dd, J=10.0, 7.5 Hz, 1 H, 7-H), 4.28 (d, J=2.5 Hz, 1 H, 10-OH), 3.54 (d, J=6.0 Hz, 1 H, 3-H), 2.88 (d, J=20.0 Hz, 1 H, 14-H), 2.75 (d, J=20.0 Hz, 1 H, 14-H), 2.54-2.47 (m, 1 H, 6-H), 2.08 (s, 3 H, Me), 2.06 (s, 3 H, Me), 1.92-1.84 (m, 1 H, 6-H), 1.77 (s, 3 H, Me), 1.31 (s, 3 H, Me), 1.15 (s, 3 H, Me), 0.88 (t, J=8.5 Hz, 9 H, Si(CH2 CH3)3), 0.55-0.45 (band, 6 H, Si(CH2 CH3)3); 13 C NMR (125 MHz, CDCl3) δ 208.4, 195.5, 170.5, 154.0, 152.0, 141.2, 88.4, 83.9, 79.8, 79.0, 76.7, 75.7, 71.9, 60.3, 43.0, 41.6, 39.8, 37.7, 31.6, 21.5, 17.8, 14.4, 9.7, 6.6, 5.0; FAB HRMS (NBA) in m/e 579.2652, M+H+ calcd for C29 H42 O10 Si 579.2626.
##STR12##
Alcohol 8. A solution of carbonate 7 (10 mg, 0.0173 mmol) in tetrahydrofuran (1 mL) at -78°C was treated with n-Butyllithium from Aldrich Chemical Company, Inc. (0.087 mL of a 1.6 M solution in hexanes, 0.139 mmol) and stirred for 1.0 hour The reaction mixture was poured into a mixture of diethylether (10 mL) and aqueous NH4 Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with diethylether (2×5 mL). The combined organic layer was washed with a saturated solution of brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 35→50% ethylacetate in hexanes) to give 8 (7.9 mg, 72%) as an amorphous solid.
Physical Data for Alcohol 8. Rf =0.36 (silica, 35% ethylacetate in petroleum ether); IR (film) νmax 3437, 2962, 2865, 1726, 1671, 1367, 1239, 1105 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 5.36 (d, J=6.5 Hz, 1H, 2-H), 5.26 (d, J=2.5 Hz, 1H, 10-H), 4.89 (br d, J=8.0 Hz, 1H, 5-H), 4.47 (d, J=8.0 Hz, 1H, 20-H), 4.32 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.26 (d, J=2.5 Hz, 1H, 10-OH), 4.15 (d, J=8.0 Hz, 1H, 20-H), 3.81 (d, J=6.5 Hz, 1H, 3-H), 2.73 (d, J=20.0 Hz, 1H, 14-H), 2.57 (d, J=20.0 Hz, 1H, 14-H), 2.49-2.41 (m, 1H, 6-H), 2.38-2.23 (m, 2H, OCCH2 (CH2)2 CH3), 2.06 (s, 3H, Me), 2.04 (s, 3H, Me), 1.90-1.82 (m, 1H, 6-H), 1.67 (s, 1H, OH), 1.64 (s, 3H, Me), 1.68-1.52 (m, 2H, OCCH2 CH2 CH2 CH3), 1.41-1.30 (m, 2H, OC(CH2)2 CH2 CH3), 1.19 (s, 3H, Me), 1.07 (s, 3 H, Me), 0.94-0.86 (band, 12H, CH3 of Bu, OSi(CH2 CH3)3), 0.58-0.45 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA) m/e 637.3421, M+H+ calcd for C33 H52 O10 Si 637.3408.
##STR13##
Alcohols 9 and 10. A solution of carbonate 7 (111.3 mg, 0.192 mmol) in tetrahydrofuran (2 mL) at -78°C was treated with vinyllithium (3.7 mL of a 0.52 M solution in diethylether, 1.92 mmol, prepared from tetravinyltin and nButyllithium: methodology from Wakefield, B. J. Organolithium Methods, Academic Press: London, 1988, p. 46) and stirred for 2.25 hour The reaction mixture was poured into a mixture of CH2 Cl2 (20 mL) and aqueous NH4 Cl (10 mL), the organic layer was separated, and the aqueous layer was extracted with CH2 Cl2 (3×10 mL). The combined organic layer was washed with brine (15 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 30→50% ethylacetate in petroleum ether) to give 9 (60.0 mg, 52%), and 10 (25.7 mg, 24%) as white foams.
Physical Data for Alcohol 9. Rf =0.52 (silica, 50% ethylacetate in hexanes); IR (film) νmax 3442, 2956, 2882, 1727, 1672, 1407, 1368, 1243, 1182, 1110, 1050, 986, 826, 736 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 6.51 (dd, J=17.0, 1.0 Hz, 1H, vinyl H), 6.13 (dd, J=17.0, 10.5 Hz, 1H, vinyl H), 6.00 (dd, J=10.5, 1.0 Hz, 1H, vinyl H), 5.45 (br d, J=6.5 Hz, 1H, 2-H), 5.30 (d, J=2.5 Hz, 1H, 10-H), 4.91 (br d, J=9.5 Hz, 1H, 5-H), 4.44 (d, J=8.5 Hz, 1H, 20-H), 4.35 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.30 (d, J=2.5 Hz, 1H, 10-OH), 4.14 (d, J=8.5 Hz, 1H, 20-H), 3.88 (d, J=6.5 Hz, 1H, 3-H), 2.79 (d, J=20.0 Hz, 1H, 14-H), 2.61 (d, J=20.0 Hz, 1H, 14-H), 2.48 (ddd, J=14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.09 (s, 3H, Me), 2.08 (s, 3H, Me), 1.89 (ddd, J=14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.72 (s, 1H, OH), 1.68 (s, 3H, Me), 1.22 (s, 3H, Me), 1.12 (s, 3H, Me), 0.92 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.62-0.46 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 739.1925, M+Cs30 calcd for C31 H46 O10 Si 739.1915.
Physical Data for Alcohol 10. Rf =0.24 (silica, 50% ethylacetate in hexanes); IR (film) νmax 3439, 2955, 2881, 1711, 1671, 1409, 1365, 1188, 1115, 980, 833, 735 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 6.48 (br d, J=17.0 Hz, 1H, vinyl H), 6.10 (dd, J=17.0, 10.5 Hz, 1H, vinyl H), 5.97 (br d, J=10.5 Hz, 1H, vinyl H), 5.47 (br d, J=6.0 Hz, 1H, 2-H), 5.25 (d, J=2.5 Hz, 1H, 10-H), 4.75 (dd, J=9.5, 3.5 Hz, 1H, 5-H), 4.38 (d, J=8.5 Hz, 1H, 20-H), 4.30 (d, J=2.5 Hz, 1H, 10-OH), 4.24 (d, J=8.5 Hz, 1H, 20-H), 3.90 (dd, J=11.5, 6.0 Hz, 1H, 7-H), 3.28 (d, J=19.5 Hz, 1H, 14-H), 3.24 (d, J=6.0 Hz, 1H, 3-H), 3.06 (br s, 1H, OH), 2.58 (d, J=19.5 Hz, 1H, 14-H), 2.38 (ddd, J=14.5, 9.5, 6.0 Hz, 1H, 6-H), 2.07 (s, 3H, Me), 1.98 (ddd, J=14.5, 11.5, 3.5 Hz, 1H, 6-H), 1.87 (s, 1H, OH), 1.61 (s, 3H, Me), 1.23 (s, 3H, Me), 1.13 (s, 3H, Me), 0.90 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.59-0.45 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 697.1802, M+Cs30 calcd for C29 H44 O9 Si 697.1809.
##STR14##
Alcohol 11. A solution of carbonate 7 (46 mg, 0.0795 mmol) in tetrahydrofuran (3 mL) at -78°C was treated with 2-furyllithium (4 mL of a 0.47 M suspension in diethylether, 1.88 mmol, prepared from furan (Aldrich Chemical Company, Inc.) and n-Butyllithium (Aldrich Chemical Company, Inc.); methodology from Ramanathan, V.; Levine, R. J. Org. Chem. 1962, 27, 1216) and stirred for 10 minutesutes The reaction mixture was poured into a mixture of CH2 Cl2 (15 mL) and aqueous NH4 Cl (20 mL). The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (10 mL), dried (MgSO4) and concentrated to give 11 which was taken into the next step without further purification.
Physical Data for Alcohol 11. Rf =0.38 (silica, 20% ethylacetate in petroleum ether); IR (film) νmax 3442, 2956, 2882, 1727, 1672, 1468, 1300, 1240, 1110, 1007, 733 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 7.66-7.64 (m, 1H, furan), 7.24 (br d, J=3.5 Hz, 1H, furan), 6.58 (dd, J=3.5, 1.5 Hz, 1H, furan), 5.55 (d, J=6.5 Hz, 1H, 2-H), 5.31 (d, J=2.0 Hz, 1H, 10-H), 4.92 (br d, J=9.0 Hz, 1H, 5-H), 4.43 (d, J=8.5 Hz, 1H, 20-H), 4.37 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.32 (d, J=2.0 Hz, 1H, 10-OH), 4.18 (d, J=8.5 Hz, 1H, 20-H), 3.93 (d, J=6.5 Hz, 1H, 3-H), 2.88 (d, J=20.0 Hz, 1H, 14-H), 2.63 (d, J=20.0 Hz, 1H, 14-H), 2.55-2.37 (m, 1H, 6-H), 2.15 (s, 3H, Me), 2.09 (s, 3H, Me), 1.93-1.87 (m, 1H, 6-H), 1.81 (s, 1H, OH), 1.71 (s, 3H, Me), 1.23 (s, 3H, Me), 1.15 (s, 3H, Me), 0.93 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.62-0.42 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/NaI) m/e 669.2717, M+Na+ calcd for C33 H46 O11 Si 669.2707.
##STR15##
Alcohol 12. A solution of carbonate 7 (50.0 mg, 0.0864 mmol) in tetrahydrofuran (5 mL) at -78°C was treated with 2-thienyllithium from Aldrich Chemical Company, inc. (1.30 mL of a 1.0 M solution in tetrahydrofuran, 1.30 mmol) and stirred for 0.5 hour The reaction mixture was poured into a mixture of diethylether (10 mL) and aqueous NH4 Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with diethylether (2×10 mL). The combined organic layer was washed with brine (10 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 10→35% ethylacetate in hexanes) to give 7 (16.5 mg, 33%), 12 (36.8 mg, 96% based on 67% conversion) as an amorphous solid.
Physical Data for Alcohol 12. Rf =0.56 (silica, 50% ethylacetate in hexanes); IR (film) νmax 3403, 2945, 2881, 1717, 1669, 1520, 1413, 1360, 1248, 1078; 1 H NMR (500 MHz, CDCl3) δ 7.84 (dd, J=3.5, 1.0 Hz, 1H, thiophene), 7.64 (d, J=1.0, 5.0 Hz, 1H, thiophene), 7.14 (dd, J=5.0, 3.5 Hz, 1H, thiophene), 5.53 (br d, J=6.5 Hz, 1H, 2-H), 5.29 (d, J=2.5 Hz, 1H, 10-H), 4.90 (br d, J=7.5 Hz, 1H, 5-H), 4.44 (d, J=8.5 Hz, 1H, 20-H), 4.35 (dd, J=10.5 Hz, 6.5 Hz, 1H, 7-H), 4.29 (d, J=2.5 Hz, 1H, 10-OH), 4.19 (d, J=8.5 Hz, 1H, 20-H), 3.90 (d, J=6.5 Hz, 1H, 3-H), 2.89 (d, J=19.5 Hz, 1H, 14-H), 2.62 (d, J=19.5 Hz, 1H, 14-H), 2.49-2.43 (m, 1H, 6-H), 2.15 (s, 3H, Me), 2.07 (s, 3H, Me), 1.92-1.84 (m, 1H, 6-H), 1.73 (s, 1H, OH), 1.71 (s, 3H, Me), 1.21 (s, 3H, Me), 1.13 (s, 3H, Me), 0.91 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.56-0.49 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA) m/e 663.2655, M+H+ calcd for C33 H46 O10 SSi 663.2659.
##STR16##
Alcohols 13 and 14. A solution of carbonate 7 (107.9 mg, 0.186 mmol) in tetrahydrofuran (6.2 mL) at -78°C was treated with 3-thienyllithium (2.76 mL of a 0.41 M solution in diethylether: tetrahydrofuran:hexanes (4.5:1:2), 1.13 mmol, prepared from 3-bromothiophene and n-Butyllithium; methodology from Camici, L.; Ricci, A.; Taddei, M. Tetrahedron Lett. 1986, 27, 5155) and stirred for 1.5 hour The reaction mixture was poured into a mixture of CH2 Cl2 (15 mL) and aqueous NH4 Cl (20 mL), the organic layer was separated, and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (10 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 20→30% ethylacetate in hexanes) to give 7 (16.9 mg, 16%), 13 (87.0 mg, 83% based on 84% conversion), and hydrolyzed C4 acetate 14 (C4-hydrolyzed side product, 9.7 mg, 10% based on 84% conversion) as amorphous solids.
Physical Data for Alcohol 13. Rf =0.74 (silica, 50% ethylacetate in hexanes), 0.41 (silica, 10% ethylacetate in benzene, 3 elutions); IR (thin film) νmax 3442, 3110, 2956, 2882, 1725, 1672, 1410, 1368, 1244, 1198, 1101, 988, 825, 744 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.18 (dd, J=3.0, 1.2 Hz, 1H, thiophene), 7.54 (dd, J=5.0, 1.2 Hz, 1H, thiophene), 7.37 (dd, J=5.0, 3.0 Hz, 1H, thiophene), 5.56 (dd, J=6.5, 1.0 Hz, 1H, 2-H), 5.31 (d, J=2.5 Hz, 1H, 10-H), 4.92 (dd, J=7.5, 2.0 Hz, 1H, 5-H), 4.40-4.34 (m, 2H, 20-H, 7-H), 4.31 (d, J=2.5 Hz, 1H, 10-OH), 4.15 (d, J=8.5 Hz, 1H, 20-H), 3.93 (d, J=6.5 Hz, 1H, 3-H), 2.88 (d, J=20 Hz, 1H, 14-H), 2.63 (dd, J=20.0, 1.0 Hz, 1H, 14-H), 2.47 (ddd, J=14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.18 (s, 3H, Me), 2.10 (s, 3H, Me), 1.89 (ddd, J=14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.81 (br s, 1H, OH), 1.72 (s, 3H, Me), 1.23 (s, 3H, Me), 1.15 (s, 3H, Me), 0.93 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.62-0.48 (band, 6H, Si(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 795.1640, M+Cs30 calcd for C33 H46 O10 SSi 795.1635.
Physical Data for Alcohol 14: Rf =0.54 (silica, 50% ethylacetate in hexanes); IR (thin film) νmax 3437, 3108, 2955, 2880, 1709, 1674, 1605, 1520, 1410, 1360, 1258, 1194, 1103, 1004, 829, 744 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.15 (dd, J=3.0, 1.0 Hz, 1H, thiophene), 7.49 (dd, J=5.0, 1.0 Hz, 1H, thiophene), 7.35 (dd, J=5.0, 3.0 Hz, 1H, thiophene), 5.59 (d, J=6.0 Hz, 1H, 2-H), 5.27 (d, J=2.5 Hz, 1H, 10-H), 4.73 (dd, J=9.5, 3.5 Hz, 1H, 5-H), 4.40 (d, J=8.5 Hz, 1H, 20-H), 4.32 (d, J=2.5 Hz, 1H, 10-OH), 4.15 (d, J=8.5 Hz, 1H, 20-H), 3.92 (dd, J=11.5, 6.0 Hz, 1H, 7-H), 3.44 (d, J=19.5 Hz, 1H, 14-H), 3.30 (d, J=6.0 Hz, 1H, 3-H), 2.91 (br s, 1H, OH), 2.61 (d, J=19.5 Hz, 1H, 14-H), 2.38 (ddd, J=14.5, 9.5, 6.0 Hz, 1H, 6-H), 2.09 (s, 3H, Me), 1.99 (ddd, J=14.5, 11.5, 3.5 Hz, 1H, 6-H), 1.81 (br s, 1H, OH), 1.65 (s, 3H, Me), 1.24 (s, 3H, Me), 1.16 (s, 3H, Me), 0.91 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.60-0.46 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 753.1530, M+Cs30 calcd for C31 H44 O9 SSi 753.1530.
##STR17##
Alcohol 15 and 16, and triol 6. A solution of carbonate 7 (62.6 mg, 0.108 mmol) in tetrahydrofuran (5.4 mL) at -78°C was treated with 2-lithiopyridine (1.15 mL of a 0.44 M solution in diethylether-pentane 1:1, 0.506 mmol, prepared from 2-bromopyridine and t-Butyllithium; methodology from Malmberg, H.; Nilsson, M. Tetrahedron, 1986, 42, 3981) and stirred for 1.3 hour The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NH4 Cl (5 mL), the organic layer was separated, and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 70→100% ethylacetate in petroleum ether) to give 6 (16.3 mg, 27%), 15 (28.0 mg, 39%), and 16 (8.4 mg, 13%) as amorphous solids.
Physical Data for Alcohol 15. Rf =0.60 (silica, ethylacetate); 1 H NMR (500 MHz, CDCl3) δ 8.77 (ddd, J=4.5, 1.7, 1.0 Hz, 1H, pyridine), 8.05 (br d, J=7.5 Hz, 1H, pyridine), 7.89 (ddd, J=7.5, 7.5, 1.7 Hz, 1H, pyridine), 7.53 (ddd, J=7.5, 4.5, 1.0 Hz, 1H, pyridine), 5.61 (dd, J=6.5, 1.0 Hz, 1H, 2-H), 5.33 (d, J=2.5 Hz, 1H, 10-H), 4.92 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.39 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.36 (d, J=9.0 Hz, 1H, 20-H), 4.33 (d, J=2.5 Hz, 1H, 10-OH), 4.28 (d, J=9.0 Hz, 1H, 20-H), 3.96 (d, J=6.5 Hz, 1H, 3-H), 2.98 (d, J=20.0 Hz, 1H, 14-H), 2.71 (dd, J=20.0, 1.0 Hz, 1H, 14-H), 2.50 (s, 1H, OH), 2.48 (ddd, J=14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.15 (s, 3H, Me), 2.11 (s, 3H, Me), 1.90 (ddd, J=14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.76 (s, 3H, Me), 1.24 (s, 3H, Me), 1.16 (s, 3H, Me), 0.93 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.63-0.47 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 790.2060, M+Cs30 calcd for C34 H47 O10 NSi 790.2024.
Physical Data for Alcohol 16. Rf =0.45 (silica, ethylacetate); IR (film) νmax 3435, 2954, 2879, 1732, 1674, 1589, 1362, 1305, 1241, 1116, 998, 829, 741 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.73 (br d, J=4.5 Hz, 1H, pyridine), 8.15 (br d, J=7.5 Hz, 1H, pyridine), 7.90 (ddd, J=7.5, 7.5, 1.7 Hz, 1H, pyridine), 7.56 (ddd, J=7.5, 4.5, 1.0 Hz, 1H, pyridine), 5.53 (dd, J=7.5, 1.0, 1H, 2-H), 5.30 (d, J=2.5 Hz, 1H, 10-H), 4.84 (dd, J=9.5, 3.0 Hz, 1H, 5-H), 4.81 (br s, 1H, OH), 4.31 (d, J=2.5 Hz, 1H, 10-OH), 4.25 (s, 2H, 20-CH2), 3.97 (dd, J=11.5, 6.5 Hz, 1H, 7-H), 3.31 (d, J=19.5 Hz, 1H, 14-H), 3.23 (d, J=7.5 Hz, 1H, 3-H), 2.57 (br d, J=19.5 Hz, 1H, 14-H), 2.43 (ddd, J=14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.11 (s, 3H, Me), 1.95 (ddd, J=14.5, 11.5, 3.0 Hz, 1H, 6-H), 1.92 (br s, 1H, OH), 1.70 (s, 3H, Me), 1.24 (s, 3H, Me), 1.17 (s, 3H, Me), 0.91 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.60-0.46 (band, 6H, OSi(CH2 CH3)3).
Physical Data for Triol 6. Rf =0.24 (silica, 50% ethylacetate in hexanes); IR (thin film) νmax 3414, 2957, 2881, 1727, 1664, 1370 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 5.23 (d, J=9.5 Hz, 1 H, 10-H), 4.89 (d, J=9.5 Hz, 1 H, 5-H), 4.63 (d, J=9.5 Hz, 1 H, 20-H), 4.56 (d, J=9.5 Hz, 1 H, 20-H), 4.32 (dd, J=11.0, 7.0 Hz, 1 H, 7-H), 4.28 (d, J=2.5 Hz, 1 H, 10-OH), 3.89 (dd, J=6.5, 4.0 Hz, 1 H, 2-H), 3.57 (d, J=6.5 Hz, 1 H, 3-H), 2.78 (d, J=19.5 Hz, 1 H, 14-H), 2.58 (d, 4.0 Hz, 1 H, 2-OH), 2.52 (d, J=19.5 Hz, 1 H, 14-H), 2.49-2.42 (m, 1 H, 6-H), 2.03 (s, 3 H, Me), 1.92-1.84 (m, 1 H, 6-H), 1.68 (s, 3 H, Me), 1.21 (s, 3 H, Me), 1.04 (s, 3 H, Me), 0.90 (t, J=8.0 Hz, 9 H, Si(CH2 CH3)3), 0.60-0.40 (band, 6 H, Si(CH2 CH3)3); 13 C NMR (125 MHz, CDCl3) δ 208.9, 198.5, 170.1, 156.7, 138.8, 83.8, 81.2, 77.6, 75.7, 72.8, 72.5, 58.8, 45.8, 43.1, 42.8, 37.3, 32.7, 21.6, 17.5, 13.6, 9.7, 6.7, 5.1; FAB HRMS (NBA/NaI) m/e 575.2648, M+Na+ calcd for C28 H44 O9 Si 575.2652.
Preparation of 3-pyridinyl-C-2 Ester Derivative (Alcohol 17) ##STR18##
Alcohol 17. To a solution of 3-lithiopyridine (1.15 mmol) in tetrahydrofuran (7 mL), prepared from 3-bromopyridine (Aldrich Chemical Company Inc.) and n-Butyllithium (Aldrich Chemical Company Inc.) at -100°C (methodology from Parham, W. E.; Piccirilli, R. M. J. Org. Chem. 1977, 42, 257), was added a solution of carbonate 7 (133.1 mg, 0.230 mmol) in tetrahydrofuran (2 mL) via cannula. The resulting solution was stirred for 1 h, allowed to warm to -78°C, stirred for 1 h, and poured into a mixture of ethylacetate (10 mL) and aqueous NH4 Cl (10 mL). The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (10 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 70→95% ethylacetate in petroleum ether) to give 7 (64.8 mg, 49%) and 17 (43.9 mg, 57% based on 51% conversion) as an amorphous solid.
Physical Data for Alcohol 17. Rf =0.56 (silica, ethylacetate); IR (film) νmax 3435, 2956, 2882, 1731, 1671, 1592, 1366, 1280, 1240, 1109, 991, 824, 739 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 9.24 (br s, 1H, pyridine), 8.81 (d, J=1.0, 4.5 Hz, 1H, pyridine), 8.30 (ddd, J=8.0, 2.0, 2.0 Hz, 1H, pyridine), 7.44 (dd, J=8.0, 4.5 Hz, 1H, pyridine), 5.66 (d, J=6.5 Hz, 1H, 2-H), 5.32 (s, 1H, 10-H), 4.92 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.38 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.32 (br s, 1H, OH), 4.30 (d, J=8.5 Hz, 1H, 20-H), 4.13 (d, J=8.5 Hz, 1H, 20-H), 3.96 (d, J=6.5 Hz, 1H, 3-H), 2.92 (d, J=19.5 Hz, 1H, 14-H), 2.66 (d, J=19.5 Hz, 1H, 14-H), 2.48 (ddd, J=15.5, 9.5, 6.5 Hz, 1H, 6-H), 2.18 (s, 3H, Me), 2.10 (s, 3H, Me), 2.03 (s, 1H, OH), 1.89 (ddd, J=14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.72 (s, 3H, Me), 1.23 (s, 3H, Me), 1.16 (s, 3H, Me), 0.92 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.62-0.48 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CSI) m/e 790.2030, M+Cs30 calcd for C34 H47 O10 NSi 790.2024.
##STR19##
Alcohol 18. A solution of carbonate 7 (150 mg, 0.259 mmol) in tetrahydrofuran (20 mL) at -78°C was treated with 4-lithio-N,N-dimethylaniline (6.5 mL of a 0.39 M solution in diethylether:pentane (3:1), 2.54 mmol, prepared from 4-bromo-N,N-dimethylaniline and t-Butyllithium; methodology from Jones, F. N.; Hauser, C. R. J. Org. Chem. 1962, 27, 4389) and stirred for 15 minutesutes The reaction mixture was poured into a mixture of CH2 Cl2 (35 mL) and aqueous NH4 Cl (20 mL), the organic layer was separated, and the aqueous layer was extracted with CH2 Cl2 (2×20 mL). The combined organic layer was washed with brine (20 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 10→35% ethylacetate in petroleum ether) to give 18 (55.0 mg, 30%) as an amorphous solid.
Physical Data for Alcohol 18. Rf =0.26 (silica, 35% ethylacetate in hexanes); IR (film) νmax 3414, 2924, 1706, 1669, 1605, 1530, 1094; 1 H NMR (500 MHz, CDCl3) δ 7.90 (d, J=9.0 Hz, 2H, Ar), 6.64 (d, J=9.0 Hz, 2H, Ar), 5.60 (br d, J=7.0 Hz, 1H, 2-H), 5.29 (d, J=2.5 Hz, 1H, 10-H), 4.89 (br d, J=9.5 Hz, 1H, 5-H), 4.37 (d, J=8.5 Hz, 1H, 20-H), 4.36 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.31 (d, J=2.5 Hz, 1H, 10-OH), 4.13 (br d, J=8.5 Hz, 1H, 20-H), 3.90 (d, J=7.0 Hz, 1H, 3-H), 3.05 (s, 6H, NMe2), 2.93 (s, 1H, OH), 2.90 (d, J=20.0 Hz, 1H, 14-H), 2.61 (br d, J=20.0 Hz, 1H, 14-H), 2.49-2.40 (m, 1H, 6-H), 2.16 (s, 3H, Me), 2.08 (s, 3H, Me), 1.90-1.83 (m, 1H, 6-H), 1.69 (s, 3H, Me), 1.20 (s, 3H, Me), 1.13 (s, 3H, Me), 0.90 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.56-0.49 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/NaI) m/e 722.3354, M+Na+ calcd for C37 H53 O10 NSi 722.3336.
##STR20##
Alcohol 19. A solution of carbonate 7 (47 mg, 0.0812 mmol) in tetrahydrofuran (2 mL) at -78°C was treated with 1-lithionaphthalene (6.3 mL of a 0.32 M solution in diethylether, 2.03 mmol, prepared from 1-bromonaphthalene from Aldrich Chemical Company Inc. and tButyllithium; methodology from Gilman, H.; Moore, F. W. J. Am. Chem. Soc. 1940, 62, 1843) and stirred for 5 minutesutes The reaction mixture was poured into a mixture of CH2 Cl2 (15 mL) and aqueous NH4 Cl (20 mL), the organic layer was separated, and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (10 mL), dried (MgSO4) and concentrated to give alcohol 19 which was taken into the next step without further purification.
Physical Data for Alcohol 19. Rf =0.27 (20% ethylacetate in petroleum ether); IR (film) νmax 3442, 2954, 2882, 1724, 1671, 1461, 1362, 1279, 1228, 1195, 1092, 987, 826, 736 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.66 (s, 1H, naphthalene), 8.07 (dd, J=9.0, 2.0 Hz, 1H, naphthalene), 7.97-7.89 (m, 3H, naphthalene), 7.68-7.57 (m, 2H, naphthalene), 5.71 (br d, J=6.5 Hz, 1H, 2-H), 5.35 (d, J=2.5 Hz, 1H, 10-H), 4.94 (br d, J=8.0 Hz, 1H, 5-H), 4.41 (dd, J=11.0, 7.0 Hz, 1H, 7-H), 4.37 (d, J=8.5 Hz, 1H, 20-H), 4.35 (d, J=2.0 Hz, 1H, 10-OH), 4.18 (d, J=8.5 Hz, 1H, 20-H), 4.00 (d, J=6.5 Hz, 1H, 3-H), 3.02 (d, J=19.5 Hz, 1H, 14-H), 2.69 (d, J=19.5 Hz, 1H, 14-H), 2.54-2.45 (m, 1H, 6-H), 2.27 (s, 3H, Me), 2.13 (s, 3H, Me), 1.94-1.87 (m, 1H, 6-H), 1.86 (s, 1H, OH), 1.75 (s, 3H, Me), 1.25 (s, 3H, Me), 1.20 (s, 3H, Me), 0.94 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.63-0.49 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA) m/e 707.3270, M+H+ calcd for C39 H50 O10 Si 707.3252.
Preparation of phenylacetylide-C-2 Ester Derivative (Alcohol 20) ##STR21##
Alcohol 20. A solution of carbonate 7 (5.0 mg, 0.00864 mmol) in tetrahydrofuran (0.5 mL) at -78°C was treated with lithium phenylacetylide from Aldrich Chemical Company Inc. (0.13 mL of a 1.0 M solution in tetrahydrofuran, 0.13 mmol) and stirred for 0.5 hour The reaction mixture was treated with aqueous NH4 Cl (0.5 mL), allowed to warm to 25°C, and diluted with H2 O (5 mL) and diethylether (5 mL). The organic layer was separated, dried, and concentrated to give a 9:1 mixture of carbonate 7 and alcohol 20 (5.0 mg, 95%) as a film.
Physical Data for Alcohol 20. Rf =0.59 (50% ethylacetate in hexanes); 1 H NMR (300 MHz, CDCl3) δ 7.63-7.57 (m, 2 H, Ar), 7.53-7.27 (m, 3 H, Ar), 5.43 (d, J=6.5 Hz, 1H, 2-H), 5.28 (d, J=2.5 Hz, 1H, 10-H), 4.90 (br d, J=7.5 Hz, 1H, 5-H), 4.67 (d, J=8.5 Hz, 1H, 20-H), 4.44 (d, J=8.5 Hz, 1H, 20-H), 4.37-4.30 (m, 1 H, 7-H), 4.28 (d, J=2.5 Hz, 1H, 10-OH), 3.88 (d, J=6.5 Hz, 1H, 3-H), 2.85 (d, J=20.2 Hz, 1H, 14-H), 2.63 (d, J=20.2 Hz, 1H, 14-H), 2.55-2.47 (m, 1 H, 6-H), 2.11 (s, 3 H, OAc), 2.08 (s, 3 H, 18-Me), 1.94-1.85 (m, 1 H, 6-H), 1.67 (s, 3 H, Me), 1.41 (s, 3 H, Me), 1.21 (s, 3 H, Me), 0.91 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.59-0.42 (band, 6H, OSi(CH2 CH3)3).
Preparation of Hydroxycarbamate-C-2 Ester Derivative (Alcohol 21) ##STR22##
Alcohol 21. A solution of carbonate 7 (5.0 mg, 0.00864 mmol) in MeOH (0.5 mL) at 25°C was treated with n-Butyl-NH2 from Aldrich Chemical Company Inc. (0.05 mL, 0.506 mmol) and stirred for 10 minutes. The reaction mixture was concentrated and purified by flash chromatography (silica, 30→50% ethylacetate in petroleum ether) to give 21 (5.2 mg, 92%) as an amorphous solid.
Physical Data for Alcohol 21. Rf =0.13 (silica, 30% ethylacetate in petroleum ether); IR (film) νmax 3434, 2957, 2881, 1711, 1671, 1368, 1243, 1108, 987, 829 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 5.27 (d, J=2.0 Hz, 1H, 10-H), 5.23 (d, J=6.5 Hz, 1H, 2-H), 4.91 (br d, J=8.0 Hz, 1H, 5-H), 4.79 (t, J=6.0 Hz, 1H, NH), 4.47 (d, J=8.5 Hz, 1H, 20-H), 4.34 (dd, J=11.0, 7.0 Hz, 1H, 7-H), 4.30 (d, J=2.5 Hz, 1H, 10-OH), 4.28 (d, J=8.5 Hz, 1H, 20-H), 3.78 (d, J=6.5 Hz, 1H, 3-H), 3.29-3.12 (m, 2H, NHCH2), 2.70 (d, J=20.0 Hz, 1H, 14-H), 2.60 (d, J=20.0 Hz, 1H, 14-H), 2.51-2.42 (m, 1H, 6-H), 2.24 (s, 1H, OH), 2.06 (s, 3H, Me), 2.05 (s, 3H, Me), 1.94-1.86 (m, 1H, 6-H), 1.69 (s, 3H, Me), 1.55-1.46 (m, 2H, NHCH2 CH2), 1.40-1.30 (m, 2H, NHCH2 CH2 CH2), 1.21 (s, 3H, Me), 1.09 (s, 3H, Me), 0.95-0.80 (m, 12H, CH3 of Bu, OSi(CH2 CH3)3), 0.61-0.47 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/NaI) m/e 674.3336, M+Na+ calcd for C33 H53 O10 NSi 674.3336.
##STR23##
Alcohol 22. A solution of carbonate 7 (5.0 mg, 0.00864 mmol) in tetrahydrofuran (0.5 mL) at -78°C was treated with LiNMePh (0.2 mL of a 0.47 M solution in diethylether, 0.094 mmol, prepared from N-methylaniline (Aldrich) and n-Butyllithium) and stirred for 1.25 hour The reaction mixture was poured into a mixture of diethylether (5 mL) and aqueous NH4 Cl (5 mL), the organic layer was separated, and the aqueous layer was extracted with diethylether (2×5 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 15→35% ethylacetate in hexanes) to give 7 (2.5 mg, 50%) and 22 (2.8 mg, 93% based on 50% conversion) as an amorphous solid.
Physical Data for Alcohol 22. Rf =0.22 (silica 35% ethylacetate in petroleum ether); 1 H NMR (500 MHz, CDCl3) δ 7.45-7.18 (band, 5H), 5.25 (br d, J=6.5 Hz, 1H, 2-H), 5.20 (d, J=2.5 Hz, 1H, 10-H), 4.70 (br d, J=8.0 Hz, 1H, 5-H), 4.26 (d, J=2.5 Hz, 1H, 10-OH), 4.22 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.19 (d, J=8.5 Hz, 1H, 20-H), 4.16 (d, J=8.5 Hz, 1H, 20-H), 3.58 (d, J=7.0 Hz, 1H, 3-H), 3.27 (s, 3H, MeN), 2.52 (d, J=20.0 Hz, 1H, 14-H), 2.35 (d, J=20.0 Hz, 1H, 14-H), 2.40-2.31 (m, 1H, 6-H), 2.03 (s, 1H, OH), 1.97 (s, 3H, Me), 1.85-1.76 (m, 1H, 6-H), 1.66 (s, 3H, Me), 1.57 (s, 3H, Me), 1.18 (s, 3H, Me), 1.08 (s, 3H, Me), 0.87 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.55-0.43 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA) m/e 686.3358, M+H+ calcd for C36 H51 O10 NSi 686.3361.
##STR24##
Alcohol 23. A solution of vinyl ester 9 (55.6 mg, 0.0916 mmol) and 4-dimethylaminutesopyridine from Aldrich Chemical Company Inc. (DMAP, 1.8 mg, 0.0147 mmol) in CH2 Cl2 (4.3 mL) at 25°C was treated with PhSH from Aldrich Chemical Company Inc. (0.030 mL, 0.292 mmol) and stirred for 1.5 hour The reaction mixture was concentrated and purified by flash chromatography (silica, 30% ethylacetate in petroleum ether) to give 23 (58.1, 88%) as a white solid.
Physical Data for Alcohol 23. Rf =0.37 (silica, 30% ethylacetate in hexanes), 0.34 (10% ethylacetate in PhH, 2 elutions); IR (film) νmax 3441, 3057, 2956, 2883, 1732, 1672, 1600, 1367, 1238, 1111, 988, 825, 739 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 7.39-7.24 (band, 5H), 5.44 (d, J=6.5 Hz, 1H, 2-H), 5.28 (d, J=2.5 Hz, 1H, 10-H), 4.90 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.38 (d, J=8.0 Hz, 1H, 20-H), 4.33 (dd, J=10.5, 6.5Hz, 1H, 7-H), 4.29 (d, J=2.5 Hz, 1H, 10-OH), 4.18 (d, J=8.0 Hz, 1H, 20-H), 3.83 (d, J=6.5 Hz, 1H, 3-H), 3.24-3.13 (m, 2H, CH2 SPh), 2.76 (d, J=19.5 Hz, 1H, 14-H), 2.72-2.58 (m, 3H, 14-H, CH2 CH2 SPh), 2.47 (ddd, J=14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.39 (s, 1H, OH), 2.07 (s, 3H, Me), 2.05 (s, 3H, Me), 1.89 (ddd, J=14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.68 (s, 3H, Me), 1.23 (s, 3H, Me), 1.12 (s, 3H, Me), 0.92 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.61-0.47 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 849.2085, M+Cs30 calcd for C37 H52 O10 SSi 849.2105.
##STR25##
Acetate 25. A solution of alcohol 11 and 4-dimethylaminopyridine (DMAP, 100 mg, 0.819 mmol) in CH2 Cl2 (3 mL) at 25°C was treated with acetic anhydride (0.50 mL, 5.30 mmol) and stirred for 3 h. The reaction mixture was diluted with CH2 Cl2 (5 mL), treated with aqueous NaHCO3 (7 mL), and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 10% ethylacetate in benzene, 3 elutions) to give 25 (36 mg, 66% from carbonate 7) as a white foam.
Physical Data for Acetate 25. Rf =0.38 (20% ethylacetate in petroleum ether); IR (film) νmax 3509, 2956, 2881, 1727, 1674, 1469, 1371, 1299, 1227, 1108, 746 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 7.65 (br s, 1H, furan), 7.24 (br d, J=3.0 Hz, 1H, furan), 6.58-6.54 (m, 2H, 10-H, furan), 5.59 (d, J=6.5 Hz, 1H, 2-H), 4.92 (br d, J=7.5 Hz, 1H, 5-H), 4.46 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.42 (d, J=8.5 Hz, 1H, 20-H), 4.16 (d, J=8.5 Hz, 1H, 20-H), 3.87 (d, J=6.5 Hz, 1H, 3-H), 2.89 (d, J=20.0 Hz, 1H, 14-H), 2.63 (d, J=20.0 Hz, 1H, 14-H), 2.59-2.48 (m, 1H, 6-H), 2.22 (s, 3H, Me), 2.17 (s, 3H, Me), 2.14 (s, 3H, Me), 1.90-1.83 (m, 1H, 6-H), 1.65 (s, 3H, Me), 1.25 (s, 3H, Me), 1.18 (s, 3H, Me), 0.91 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.64-0.52 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 821.1966, M+Cs30 calcd for C35 H48 O12 Si 821.1969.
Alcohol 26. A solution of enone 25 (36 mg, 0.0523 mmol) in MeOH (3 mL) containing two drops of CH3 COOH at 0°C was treated with NaBH4 (200 mg, 5.29 mmol, added by portions) and stirred for 6 h. The reaction mixture was diluted with CH2 Cl2 (10 mL), treated with aqueous NH4 Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 50% ethylacetate in petroleum ether) to give 26 (30 mg, 83%) as an amorphous solid.
Physical Data for Alcohol 26. Rf =0.42 (silica, 50% ethylacetate in petroleum ether); 1 H NMR (300 MHz, CDCl3) δ 7.62 (br s, 1H, furan), 7.25 (d, J=3.5 Hz, 1H, furan), 6.58 (d, J=3.5 Hz, 1H, furan), 6.43 (s, 1 H, 10-H), 5.51 (d, J=7.0 Hz, 1H, 2-H), 4.96 (d, J=7.5 Hz, 1H, 5-H), 4.85-4.79 (m, 1 H, 13-H), 4.48 (dd, J=10.5, 7.5 Hz, 1H, 7-H), 4.38 (d, J=8.0 Hz, 1H, 20-H), 4.15 (d, J=8.0 Hz, 1H, 20-H), 3.82 (d, J=7.0 Hz, 1H, 3-H), 2.61-2.48 (m, 2 H, 6-H and 14-H), 2.28 (s, 3 H, OAc), 2.20-2.10 (m, 1 H, 14-H), 2.18 (s, 6 H, OAc and 18-Me), 1.98-1.80 (m, 1 H, 6-H), 1.18 (s, 3 H, 16-Me), 1.04 (s, 3 H, 17-Me), 0.90 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.65-0.50 (band, 6H, OSi(CH2 CH3)3).
DiTES taxoid 27. To a solution of alcohol 26 (30.0 mg, 0.0434 mmol, previously azeotroped twice with benzene) and β-lactam 24 (28.0 mg, 0.0734 mmol, previously azeotroped twice with benzene) in THF (2 mL), prepared from the Ojima-Holton protocol (Holton, R. A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M.; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M.; Zucco, M.; Park, Y. H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), at 0°C was added NaN(SiMe3)2 (0.130 mL of a 1.0 M solution in THF, 0.130 mmol) dropwise. The resulting solution was stirred for 5 min and poured into a mixture of CH2 Cl2 (10 mL) and aqueous NH4 Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×5 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 60% ethylacetate in petroleum ether) to give 27 (12 mg, 26%) as an amorphous solid which was taken directly into the next step.
Taxoid 28. A solution of silyl ether 27 (6 mg, 0.00560 mmol) in THF (1 mL) at 25°C was treated with HF•-pyridine (1 mL) and stirred for 1 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHCO3 (10 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 60% ethylacetate in petroleum ether) to give 28 (3 mg, 64%) as a colorless film.
Physical Data for Taxoid 28. Rf =0.1 (50% ethylacetate in petroleum ether); IR (film) νmax 3383, 2933, 2898, 1729, 1649, 1519, 1242, 1110, 1071 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 7.77-7.73 (m, 2H), 7.68-7.66 (m, 1H, furan), 7.55-7.33 (band, 9H), 6.98 (d, J=9.0 Hz, 1H, NH), 6.58 (dd, J=3.5, 1.5 Hz, 1H, furan), 6.27-6.21 (m, 2H, 10-H, 13-H), 5.80 (dd, J=9.0, 2.0 Hz, 1H, 3'-H), 5.57 (d, J=7.0 Hz, 1H, 2-H), 4.96 (dd, J=10.0, 2.0 Hz, 1H, 5-H), 4.80 (d, J=2.0 Hz, 1H, 2'-H), 4.43-4.37 (m, 2H, 7-H, 20-H), 4.24 (d, J=8.5 Hz, 1H, 20-H), 3.77 (d, J=7.0 Hz,1 H, 3-H), 2.60-2.52 (m, 1H, 6-H), 2.47 (d, J=4.0 Hz, 1H, OH), 2.38 (s, 3H, Me), 2.35-2.21 (m, 2H, 14-CH2), 2.25 (s, 3H, Me), 1.94-1.86 (m, 1H, 6-H), 1.81 (br s, 3 H, Me), 1.76 (s, 1H, OH), 1.68 (s, 3H, Me), 1.25 (s, 3H, Me), 1.13 (s, 3H, Me).
##STR26##
Acetate 29. A solution of alcohol 12 (36.0 mg, 0.0543 mmol) and 4-dimethylaminopyridine (DMAP, 33.0 mg, 0.270 mmol) in CH2 Cl2 (3.0 mL) at 25°C was treated with acetic anhydride (0.50 mL, 5.30 mmol) and stirred for 1 h. The reaction mixture was diluted with CH2 Cl2 (10 mL), treated with aqueous NaHCO3 (7 mL), and stirred vigorously for 0.5 h. The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 10→35% ethylacetate in hexanes) to give 29 (29.5 mg, 77%) as an amorphous solid.
Physical Data for Acetate 29. Rf =0.56 (silica, 50% ethylacetate in petroleum ether); IR (film) νmax 3457, 2956, 1712, 1669, 1525, 1413, 1376, 1264, 1227, 1073; 1 H NMR (500 MHz, CDCl3) δ 7.84 (dd, J=4.0, 1.5 Hz, 1H, thiophene), 7.63 (dd, J=5.0, 1.5 Hz, 1H, thiophene), 7.13 (dd, J=5.0, 4.0 Hz, 1H, thiophene), 6.56 (s, 1H, 10-H), 5.58 (br d, J=6.5 Hz, 1H, 2-H), 4.90 (br d, J=8.0 Hz, 1H, 5-H), 4.44 (dd, J=10.5, 7.0 Hz, 1H, 7-H), 4.42 (d, J=8.5 Hz, 1H, 20-H), 4.18 (d, J=8.5 Hz, 1H, 20-H), 3.85 (d, J=6.5 Hz, 1H, 3-H), 2.91 (d, J=19.5 Hz, 1H, 14-H), 2.64 (dd, J=19.5, 1.0 Hz, 1H, 14-H), 2.55-2.48 (m, 1H, 6-H), 2.20 (s, 3H, Me), 2.15 (s, 3H, Me), 2.14 (s, 3H, Me), 1.89-1.82 (m, 1H, 6-H), 1.65 (s, 3H, Me), 1.23 (s, 3H, Me), 1.16 (s, 3H, Me), 0.88 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.59-0.53 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 837.1736, M+Cs30 calcd for C35 H48 O10 SSi 837.1741.
Alcohol 30. A solution of enone 29 (29.0 mg, 0.0411 mmol) in MeOH (5 mL) at 0°C was treated with NaBH4 (30.2 mg, 0.80 mmol, added by portions) and stirred for 2.5 h. The reaction mixture was diluted with CH2 Cl2 (15 mL), treated with aqueous NH4 Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 25→50% ethylacetate in petroleum ether) to give 29 (4.0 mg, 14%) and 30 (14.7 mg, 59% based on 86% conversion) as an amorphous solid.
Physical Data for Alcohol 30. Rf =0.34 (silica, 50% ethylacetate in petroleum ether); IR (film) νmax 3478, 2946, 2892, 1717, 1520, 1365, 1238, 1083; 1 H NMR (500 MHz, CDCl3) δ 7.85 (dd, J=3.5, 1.5 Hz, 1H, thiophene), 7.61 (dd, J=5.0, 1.5 Hz, 1H, thiophene), 7.12 (dd, J=5.0, 3.5 Hz, 1H, thiophene), 6.43 (s, 1H, 10-H), 5.51 (d, J=7.0 Hz, 1H, 2-H), 4.94 (br d, J=7.5 Hz, 1H, 5-H), 4.83-4.77 (m, 1H, 13-H), 4.45 (dd, J=10.5, 7.5 Hz, 1H, 7-H), 4.41 (d, J=8.0 Hz, 1H, 20-H), 4.19 (br d, J=8.0 Hz, 1H, 20-H), 3.82 (d, J=7.0 Hz, 1H, 3-H), 2.55-2.48 (m, 1H, 6-H), 2.24 (s, 3H, Me), 2.26-2.21 (m, 2H, 14-CH2), 2.16 (d, J=1.0 Hz, 3H, 18-Me), 2.15 (s, 3H, Me), 2.00 (d, J=5.0 Hz, 1H, OH), 1.90-1.82 (m, 1H, 6-H), 1.66 (s, 3H, Me), 1.58 (s, 1H, OH), 1.15 (s, 3H, Me), 1.02 (s, 3H, Me), 0.90 (t, J=8.0 Hz, 9H, OSiCH2 CH3)3), 0.59-0.55 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 839.1893, M+Cs30 calcd for C35 H50 O11 SSi 839.1897.
DiTES taxoid 31. To a solution of alcohol 30 (14.5 mg, 0.0205 mmol, previously azeotroped twice with benzene) and β-lactam 24 (16.0 mg, 0.0420 mmol, previously azeotroped twice with benzene) in THF (1.0 mL), prepared from the Ojima-Holton protocol (Holton, R. A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.;
Zhao, M.; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M.; Zucco, M.; Park, Y. H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), at -78°C was added NaN(SiMe3)2 (0.051 mL of a 1.0 M solution in THF, 0.051 mmol) dropwise. The resulting solution was stirred for 0.5 h and poured into a mixture of diethylether (10 mL) and aqueous NH4 Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with diethylether (2×5 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 10→35% ethylacetate in hexanes) followed by preparative TLC (silica, 15% ethylacetate in benzene) to give 30 (3.0 mg, 21%) and 31 (7.6 mg, 43% based on 79% conversion) as a white solid.
Physical Data for DiTES taxoid 31. Rf =0.48 (silica, 50% ethylacetate in hexanes); IR (film) νmax 3382, 2913, 2850, 1722, 1653, 1461, 1243, 1083, 1014; 1 H NMR (500 MHz, CDCl3) δ 7.90 (br d, J=4.0 Hz, 1H, thiophene), 7.74 (d, J=8.0 Hz, 2H, NBz), 7.62 (br d, J=5.0 Hz, 1H, thiophene), 7.48 (t, J=7.0 Hz, 1H, Ar), 7.42-7.28 (band, 7H, Ar), 7.14 (dd, J=5.0, 4.0 Hz, 1H, thiophene), 7.10 (d, J=9.0 Hz, 1H, NH), 6.42 (s, 1H, 10-H), 6.20 (br t, J=9.0 Hz, 1H, 13-H), 5.65 (br d, J=9.0 Hz, 1H, 3'-H), 5.57 (d, J=7.0 Hz, 1H, 2-H), 4.94 (br d, J=8.5 Hz, 1H, 5-H), 4.67 (d, J=1.5 Hz, 1H, 2'-H), 4.44 (dd, J=11.0, 6.5 Hz, 1H, 7-H), 4.43 (d, J=8.5 Hz, 1H, 20-H), 4.26 (d, J=8.5 Hz, 1H, 20-H), 3.77 (d, J=7.0 Hz, 1H, 3-H), 2.51 (s, 3H, Me), 2.54-2.47 (m, 1H, 6-H), 2.34 (dd, J=15.0, 9.5 Hz, 1H, 14-H), 2.15 (s, 3H, Me), 2.10 (dd, J=15.0, 9.0, 1H, 14-H), 1.99 (s, 3H, Me), 1.93-1.86 (m, 1H, 6-H), 1.72 (s, 1H, OH), 1.68 (s, 3H, Me), 1.18 (s, 3H, Me), 1.16 (s, 3H, Me), 0.90 (t, J=8.0 Hz, 9H, Si(CH2 CH3)3), 0.79 (t, J=8.0 Hz, 9H, Si(CH2 CH3)3), 0.57-0.55 (band, 6H, Si(CH2 CH3)3), 0.45-0.40 (band, 6H, Si(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 1220.3685 M+Cs30 calcd for C57 H77 O14 NSSi2 1220.3658.
Taxoid 32. A solution of silyl ether 31 (7.5 mg, 0.00689 mmol) in THF (0.8 mL) at 25°C was treated with HF•-pyridine (0.150 mL) and stirred for 1 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHCO3 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 50→100% ethylacetate in petroleum ether) to give 32 (4.2 mg, 71%) as a colorless film.
Physical Data for Taxoid 32. Rf =0.44 (silica, 75% ethylacetate in petroleum ether); IR (film) νmax 3417, 2929, 1716, 1649, 1521, 1460, 1417, 1368, 1247, 1076; 1 H NMR (500 MHz, CDCl3) δ 7.90 (dd, J=4.0, 1.0 Hz, 1H, thiophene), 7.73 (d, J=7.0 Hz, 2H, NBz), 7.63 (dd, J=5.0, 1.0 Hz, 1H, thiophene), 7.51-7.32 (band, 8H, Ar), 7.14 (dd, J=5.0, 4.0 Hz, 1H, thiophene), 6.96 (d, J=9.0 Hz, 1H, NH), 6.24 (s, 1H, 10-H), 6.19 (br t, J=9.0 Hz, 1H, 13-H), 5.75 (dd, J=9.0, 2.5 Hz, 1H, 3'-H), 5.55 (d, J=7.0 Hz, 1H, 2-H), 4.94 (br d, J=8.0 Hz, 1H, 5-H), 4.76 (dd, J=5.0, 2.5 Hz, 1H, 2'-H), 4.41 (d, J=8.5 Hz, 1H, 20-H), 4.40-4.33 (m, 1H, 7-H), 4.24 (d, J=8.5 Hz, 1H, 20-H), 3.73 (d, J=7.0 Hz, 1H, 3-H), 3.52 (d, J=5.0 Hz, 1H, 2'-OH), 2.58-2.49 (m, 1H, 6-H), 2.44 (d, J=4.0 Hz, 1H, 7-OH), 2.35 (s, 3H, Me), 2.29 (d, J=9.0 Hz, 2H, 14-CH2), 2.22 (s, 3H, Me), 1.91-1.83 (m, 1H, 6-H), 1.76 (s, 3H, Me), 1.66 (s, 3H, Me), 1.23 (s, 3H, Me), 1.10 (s, 3H, Me); FAB HRMS (NBA/CsI) m/e 992.1252, M+Cs30 calcd for C45 H49 NO14 S 992.1928.
##STR27##
Acetate 33. A solution of alcohol 13 (68.4 mg, 0.103 mmol) and 4-dimethylaminopyridine (DMAP, 37.8 mg, 0.309 mmol) in CH2 Cl2 (4.4 mL) at 25°C was treated with acetic anhydride (0.370 mL, 3.92 mmol) and stirred for 2 h. The reaction mixture was diluted with CH2 Cl2 (5 mL), treated with aqueous NaHCO3 (7 mL), and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 30% ethylacetate in hexanes) to give 33 (66.0 mg, 91%) as an amorphous solid.
Physical Data for Acetate 33. Rf =0.48 (silica, 10% ethylacetate in benzene, 3 elutions); IR (film) νmax 3518, 2956, 2881, 1727, 1676, 1520, 1460, 1371, 1236, 1098, 985, 824, 744 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.19 (dd, J=3.0, 1.1 Hz, 1H, thiophene), 7.55 (dd, J=5.0, 1.1 Hz, 1H, thiophene), 7.38 (dd, J=5.0, 3.0 Hz, 1H, thiophene), 6.58 (s, 1H, 10-H), 5.61 (dd, J=6.5, 0.7 Hz, 1H, 2-H), 4.92 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.47 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.38 (d, J=8.5 Hz, 1H, 20-H), 4.14 (d, J=8.5 Hz, 1H, 20-H), 3.88 (d, J=6.5 Hz, 1H, 3-H), 2.89 (d, J=20 Hz, 1H, 14-H), 2.64 (br d, J=20 Hz, 1H, 14-H), 2.54 (ddd, J=14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.23 (s, 3H, Me), 2.18 (s, 3H, Me), 2.17 (s, 3H, Me), 1.87 (ddd, J=14.5, 10.5, 2.0, 1H, 6-H), 1.85 (s, 1H, OH), 1.66 (s, 3H, Me), 1.26 (s, 3H, Me), 1.19 (s, 3H, Me), 0.92 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.65-0.54 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 837.1760, M+Cs30 calcd for C35 H48 O11 SSi 837.1741.
Alcohol 34. A solution of enone 33 (57.3 mg, 0.0813 mmol) in MeOH-THF (5:1, 4.1 mL) at 0°C was treated with NaBH4 (69.1 mg, 1.83 mmol, added by portions) and stirred for 2.5 h. The reaction mixture was diluted with CH2 Cl2 (10 mL), treated with aqueous NH4 Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 30% ethylacetate in hexanes) to give 33 (6.8 mg, 12%) and 34 (45.2 mg, 89% based on 88% conversion) as an amorphous solid.
Physical Data for Alcohol 34. Rf =0.48 (silica, 50% ethylacetate in hexanes); IR (film) νmax 3520, 2953, 2881, 1719, 1520, 1370, 1238, 1100, 979, 823, 746 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.20 (dd, J=3.0, 1.0 Hz, 1H, thiophene), 7.57 (dd, J=5.0, 1.0 Hz, 1H, thiophene), 7.35 (dd, J=5.0, 3.0 Hz, 1H, thiophene), 6.45 (s, 1H, 10-H), 5.54 (d, J=7.0 Hz, 1H, 2-H), 4.96 (br d, J=8.5 Hz, 1H, 5-H), 4.82 (br dd, J=12.0, 8.0 Hz, 1H, 13-H), 4.48 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.36 (d, J=8.5 Hz, 1H, 20-H), 4.15 (d, J=8.5 Hz, 1H, 20-H), 3.85 (d, J=7.0 Hz, 1H, 3-H), 2.53 (ddd, J=14.5, 9.5, 6.5, 1H, 6-H), 2.27 (s, 3H, Me), 2.28-2.21 (m, 2H, 14-CH2), 2.18 (s, 6H, Me, Me), 2.03 (s, 1H, OH), 1.87 (ddd, J=14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.67 (s, 3H, Me), 1.65 (s, 1H, OH), 1.18 (s, 3H, Me), 1.04 (s, 3H, Me), 0.92 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.64-0.50 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 839.1908 M+Cs30 calcd for C35 H50 O11 SSi 839.1897.
DiTES taxoid 35. To a solution of alcohol 34 (19.5 mg, 0.0276 mmol, previously azeotroped twice with benzene) and β-lactam 24 (27.5 mg, 0.0721 mmol, previously azeotroped twice with benzene) in THF (1.4 mL), prepared from the Ojima-Holton protocol (Holton, R. A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M.; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M.; Zucco, M.; Park, Y. H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), at 0°C was added NaN(SiMe3)2 (0.066 mL of a 1.0 M solution in THF, 0.066 mmol) dropwise. The resulting solution was stirred for 0.5 h and poured into a mixture of CH2 Cl2 (10 mL) and aqueous NH4 Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×5 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 20→30% ethylacetate in hexanes) to give 34 (1.1 mg, 6%) and 35 (17.3 mg, 61% based on 94% conversion) as a white solid.
Physical Data for DiTES taxoid 35. Rf =0.86 (silica, 50% ethylacetate in hexanes); IR (film) νmax 3519, 3437, 2953, 2879, 1726, 1666, 1515, 1483, 1369, 1240, 1100, 979, 825, 746 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.32 (dd, J=3.0, 1.2 Hz, 1H, thiophene), 7.76-7.73 (m, 2H), 7.60 (dd, J=5.0, 1.2 Hz, 1H, thiophene), 7.52-7.29 (band, 9H), 7.10 (d, J=9.0 Hz, 1H, NH), 6.44 (s, 1H, 10-H), 6.26 (br t, J=9.0 Hz, 1H, 13-H), 5.72 (dd, J=9.0, 2.0 Hz, 1H, 3'-H), 5.61 (d, J=7.0 Hz, 1H, 2-H), 4.95 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.70 (d, J=2.0 Hz, 1H, 2'-H), 4.48 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.37 (d, J=8.5 Hz, 1H, 20-H), 4.23 (d, J=8.5 Hz, 1H, 20-H), 3.81 (d, J=7.0 Hz, 1H, 3-H), 2.56-2.49 (m, 1H, 6-H), 2.54 (s, 3H, Me), 2.35 (dd, J=15.5, 9.0 Hz, 1H, 14-H), 2.17 (s, 3H, Me), 2.07 (dd, J=15.5, 9.0 Hz, 1H, 14-H), 2.03 (d, J=1.0 Hz, 3H, 18-Me), 1.94-1.87 (m, 1H, 6-H), 1.69 (s, 3H, Me), 1.68 (s, 1H, OH), 1.20 (s, 3H, Me), 1.18 (s, 3H, Me), 0.93 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.81 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.63-0.53 (band, 6H, OSi(CH2 CH3)3), 0.52-0.36 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 1220.3675, M+Cs30 calcd for C57 H77 O14 SSi2 N 1220.3658.
Taxoid 36. A solution of silyl ether 35 (17.3 mg, 0.0159 mmol) in THF (0.6 mL) at 25°C was treated with HF•pyridine (0.150 mL) and stirred for 2 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHCO3 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 25% ethylacetate In petroleum ether) to give 36 (7.7 mg, 56%) as a colorless film.
Preparation of Taxoid 36. Rf =0.11 (silica, 50% ethylacetate in hexanes); IR (film) νmax 3496, 3434, 2940, 1723, 1648, 1519, 1370, 1243, 1071, 975 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.32 (dd, J=3.0, 1.0 Hz, 1H, thiophene), 7.75-7.72 (m, 2H), 7.60 (dd, J=5.0, 1.0 Hz, 1H, thiophene), 7.53-7.33 (band, 9H), 6.95 (d, J=9.0 Hz, 1H, NH), 6.28-6.23 (m, 2H, 10-H, 13-H), 5.81 (dd, J=9.0, 2.0 Hz, 1H, 3'-H), 5.58 (d, J=7.0 Hz, 1H, 2-H), 4.95 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.80 (dd, J=4.5, 2.0 Hz, 1H, 2'-H), 4.41 (br t, J=7.5 Hz, 1H, 7-H), 4.36 (d, J=8.5 Hz, 1H, 20-H), 4.22 (d, J=8.5 Hz, 1H, 20-H), 3.78 (d, J=7.0 Hz, 1H, 3-H), 3.49 (d, J=4.5 Hz, 1H, 2'-OH), 2.55 (ddd, J=14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.45 (br s, 1H, OH), 2.40 (s, 3H, Me), 2.34 (dd, J=15.5, 9.0 Hz, 1H, 14-H), 2.25 (dd, J=15.5, 9.0 Hz, 1H, 14-H), 2.24 (s, 3H, Me), 1.89 (ddd, J=14.5, 11.0, 2.0 Hz, 1H, 6-H), 1.81 (d, J=2.0 Hz, 3H, 18-Me), 1.74 (br s, 1H, OH), 1.67 (s, 3H, Me), 1.24 (s, 3H, Me), 1.13 (s, 3H, Me); FAB HRMS (NBA/CsI) m/e 992.1940, M+Cs30 calcd for C45 H49 O14 NS 992.1928.
##STR28##
Acetate 37. A solution of alcohol 15 (23.2 mg, 0.0353 mmol) and 4-dimethylaminopyridine (DMAP, 12.9 mg, 0.106 mmol) in CH2 Cl2 (1.5 mL) at 25°C was treated with acetic anhydride (0.126 mL, 1.34 mmol) and stirred for 2 h. The reaction mixture was diluted with ethylacetate (5 mL), treated with aqueous NaHCO3 (7 mL), and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 70→100% ethylacetate in petroleum ether) to give 37 (19.0 mg, 77%) as an amorphous solid.
Preparation of Acetate 37. Rf =0.58 (silica, ethylacetate); IR (film) νmax 3482, 2954, 2881, 1730, 1675, 1370, 1304, 1231, 1118, 987, 823, 739 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.77 (ddd, J=4.5, 1.7, 1.0 Hz, 1H, pyridine), 8.05 (ddd, J=8.0, 1.0, 1.0 Hz, 1H, pyridine), 7.89 (ddd, J=8.0, 8.0, 1.7 Hz, 1H, pyridine), 7.53 (ddd, J=8.0, 4.5, 1.0 Hz, 1H, pyridine), 6.59 (s, 1H, 10-H), 5.65 (dd, J=6.6, 1.0 Hz, 1H, 2-H), 4.92 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.48 (dd, J=10.5, 7.0 Hz, 1H, 7-H), 4.35 (d, J=8.5 Hz, 1H, 20-H), 4.26 (dd, J=8.5, 1.0 Hz, 1H, 20-H), 3.91 (d, J=6.5 Hz, 1H, 3-H), 3.00 (d, J=20.0 Hz, 1H, 14-H), 2.71 (dd, J=20.0, 1.0 Hz, 1H, 14-H), 2.54 (ddd, J=14.5, 9.5, 7.0 Hz, 1H, 6-H), 2.53 (s, 1H, OH), 2.23 (s, 3H, Me), 2.18 (s, 3H, Me), 2.14 (s, 3H, Me), 1.88 (ddd, J=14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.70 (s, 3H, Me), 1.27 (s, 3H, Me), 1.20 (s, 3H, Me), 0.92 (t, 9H, J=8.0 Hz, OSi(CH2 CH3)3), 0.64-0.52 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 832.2139, M+Cs30 calcd for C36 H49 O11 NSi 832.2129.
Alcohol 38. A solution of enone 37 (47.6 mg, 0.0680 mmol) in MeOH-THF (5:1, 3.8 mL) at 0°C was treated with NaBH4 (46.0 mg, 1.22 mmol, added by portions) and stirred for 1.5 h. The reaction mixture was diluted with ethylacetate (10 mL), treated with aqueous NH4 Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (basic alumina, ethylacetate→10% MeOH in ethylacetate) to give 27f (28.0 mg, 59%) as an amorphous solid.
Physical Data for Alcohol 38. Rf =0.36 (silica, ethylacetate); IR (film) νmax 3487, 2951, 2880, 1736, 1583, 1369, 1307, 1236, 1132, 983, 824, 739 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.79 (dm, J=4.5 Hz, 1H, pyridine), 8.13 (br d, J=7.5 Hz, 1H, pyridine), 7.88 (ddd, J=7.5, 7.5, 1.7 Hz, 1H, pyridine), 7.51 (ddd, J=7.5, 4.5, 1.0 Hz, 1H, pyridine), 6.46 (s, 1H, 10-H), 5.64 (d, J=7.0 Hz, 1H, 2-H), 4.96 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.85 (br t, J=8.0 Hz, 1H, 13-H), 4.49 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.31 (d, J=8.0 Hz, 1H, 20-H), 4.25 (d, J=8.0 Hz, 1H, 20-H), 3.89 (d, J=7.0 Hz, 1H, 3-H), 2.53 (ddd, J=14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.36-2.11 (m, 2H, 14-CH2), 2.25 (s, 3H, Me), 2.19 (d, J=1.0 Hz, 3H, 18-Me), 2.18 (s, 3H, Me), 1.88 (ddd, J=14.0, 10.5, 2.5 Hz, 1H, 6-H), 1.70 (s, 3H, Me), 1.20 (s, 3H, Me), 1.05 (s, 3H, Me), 0.92 (t, J=8.0 Hz, 9H, OSi(CH2 CH33), 0.65-0.51 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 834.231 1, M+Cs30 calcd for C36 H51 O11 NSi 834.2286.
DiTES taxoid 39. To a solution of alcohol 38 (10.3 mg, 0.0147 mmol, previously azeotroped twice with benzene) and β-lactam 24 (17.0 mg, 0.0446 mmol, previously azeotroped twice with benzene) in THF (0.75 mL) at 0°C, prepared from the Ojima-Holton protocol (Holton, R. A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M.; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M.; Zucco, M.; Park, Y. H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), was added NaN(SiMe3)2 (0.038 mL of a 1.0 M solution in THF, 0.038 mmol) dropwise. The resulting solution was stirred for 20 min and poured into a mixture of ethylacetate (10 mL) and aqueous NH4 Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×5 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 60% ethylacetate in petroleum ether) to give 39 (2.7 mg, 17%) as a film.
Physical Data for DiTES taxoid 39. Rf =0.28 (silica, 50% ethylacetate in hexane); IR (film) νmax 3429, 2952, 2927, 2878, 1728, 1662, 1585, 1369, 1236, 1124, 1016, 984, 742 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.78 (br d, J=4.5 Hz, 1H, pyridine), 8.21 (d, J=8.0 Hz, 1H, pyridine), 7.95 (ddd, J=8.0, 8.0, 1.7 Hz, 1H, pyridine), 7.75-7.70 (m, 2H), 7.54-7.22 (band, 9H), 7.12 (d, J=9.0 Hz, 1H, NH), 6.45 (s, 1H, 10-H), 6.27 (br t, J=9.0 Hz, 1H, 13-H), 5.73-5.67 (m, 2H, 2-H, 3'-H), 4.95 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.70 (d, J=2.0 Hz, 1H, 2'-H), 4.48 (dd, J=10.5, 6.5, 1H, 7-H), 4.32 (br s, 2H, 20-CH2), 3.85 (d, J=7.0 Hz, 1H, 3-H), 2.56-2.48 (m, 1H, 6-H), 2.52 (s, 3H, Me), 2.40 (dd, J=15.0 Hz, 9.5 Hz, 1H, 14-H), 2.20-2.12 (m, 2H, 14-H, OH), 2.18 (s, 3H, Me), 2.04 (s, 3H, Me), 1.92 (ddd, J=14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.72 (s, 3H, Me), 1.22 (s, 3H, Me), 1.19 (s, 3H, Me), 0.93 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.81 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.64-0.34 (band, 12H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 1215.4065, M+Cs30 calcd for C58 H78 O14 N2 Si2 1215.4046.
Taxoid 40. A solution of silyl ether 39 (2.7 mg, 0.00249 mmol) in THF (0.4 mL) at 25°C was treated with HF•pyridine (0.170 mL) and stirred for 3 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHCO3 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, ethylacetate) to give 40 (0.8 mg, 38%) as a colorless film.
Physical Data for Taxoid 40. Rf =0.54 (silica, ethylacetate); 1 H NMR (500 MHz, CDCl3) δ 8.80 (br d, J=4.5 Hz, 1H, pyridine), 8.22 (d, J=7.5 Hz, 1H, pyridine), 7.93 (ddd, J=7.5, 7.5, 1.5 Hz, 1H, pyridine), 7.75-7.71 (m, 2H), 7.54-7.30 (band, 9H), 6.98 (d, J=9.0 Hz, 1H, NH), 6.30-6.24 (m, 2H, 10-H, 13-H), 5.82 (dd, J=9.0, 2.5 Hz, 1H, 3'-H), 5.67 (d, J=7.0 Hz, 1H, 2-H), 4.95 (dd, J=10.0, 2.0 Hz, 1H, 5-H), 4.81 (dd, J=4.5, 2.5 Hz, 1H, 2'-H), 4.41 (ddd, J=11.0, 7.0, 4.5 Hz, 1H, 7-H), 4.31 (s, 2H, 20-CH2), 3.81 (d, J=7.0 Hz, 1H, 3-H), 3.52 (br s, 1H, OH), 3.50 (d, J=4.5 Hz, 1H, 2'-OH), 2.56 (ddd, J=14.5, 9.5, 7.0 Hz, 1H, 6-H), 2.46 (d, J=4.0 Hz, 1H, 7-OH), 2.43-2.30 (m, 2H, 14-CH2), 2.38 (s, 3H, OAc), 2.25 (s, 3H, OAc), 1.90 (ddd, J=14.5, 11.0, 2.0 Hz, 1H, 6-H), 1.81 (s, 3H, Me), 1.71 (s, 3H, Me), 1.26 (s, 3H, Me), 1.15 (s, 3H, Me).
##STR29##
Acetate 41. A solution of alcohol 17 (42.9 mg, 0.0652 mmol) and 4-dimethylaminopyridine (DMAP, 23.9 mg, 0.196 mmol) in CH2 Cl2 (2.8 mL) at 25°C was treated with acetic anhydride (0.235 mL, 2.49 mmol) and stirred for 2 h. The reaction mixture was diluted with ethylacetate (5 mL), treated with aqueous NaHCO3 (7 mL), and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, ethylacetate) to give 41 (43.5 mg, 95%) as a white solid.
Physical Data for Acetate 41. Rf =0.61 (silica, ethylacetate); IR (film) νmax 3470, 3327, 2955, 2881, 1731, 1675, 1592, 1370, 1279, 1229, 1108, 822, 738 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 9.23 (br s, 1H, pyridine), 8.79 (br s, 1H, pyridine), 8.30 (ddd, J=8.0, 2.0, 2.0 Hz, 1H, pyridine), 7.43 (dd, J=8.0, 5.0 Hz, 1H, pyridine), 6.58 (s, 1H, 10-H), 5.70 (dd, J=6.5, 1.0 Hz, 1H, 2-H), 4.91 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.47 (dd, J=10.5, 7.0 Hz, 1H, 7-H), 4.28 (d, J=8.0 Hz, 1H, 20-H), 4.11 (d, J=8.0 Hz, 1H, 20-H), 3.91 (d, J=6.5 Hz, 1H, 3-H), 2.93 (d, J=20.0 Hz, 1H, 14-H), 2.68 (dd, J=20.0, 1.0 Hz, 1H, 14-H), 2.53 (ddd, J=14.5, 9.5, 7.0 Hz, 1H, 6-H), 2.24 (br s, 1H, OH), 2.22 (s, 3H, Me), 2.18 (s, 3H, Me), 2.17 (s, 3H, Me), 1.85 (ddd, J=14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.66 (s, 3H, Me), 1.26 (s, 3H, Me), 1.18 (s, 3H, Me), 0.90 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.63-0.51 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 832.2145, M+Cs30 calcd for C36 H49 O11 NSi 832.2129.
Alcohol 42. A solution of enone 41 (39.8 mg, 0.0569 mmol) in MeOH-THF (5:1, 3.1 mL) at 0°C was treated with NaBH4 (65.0 mg, 1.72 mmol, added by portions) and stirred for 1.5 h. The reaction mixture was diluted with ethylacetate (10 mL), treated with aqueous NH4 Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, ethylacetate) to give 41 (3.7 mg, 9%) and 42 (24.3 mig, 67% based on 91% conversion) as an amorphous solid.
Physical Data for Alcohol 42. Rf =0.42 (silica, ethylacetate); IR (film) νmax 3490, 2953, 2881, 1727, 1592, 1369, 1235, 1110, 822, 740 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 9.30 (d, J=2.0 Hz, 1H, pyridine), 8.81 (dd, J=5.0, 2.0 Hz, 1H, pyridine), 8.35 (ddd, J=8.0, 2.0, 2.0 Hz, 1H, pyridine), 7.44 (dd, J=8.0, 5.0 Hz, 1H, pyridine), 6.46 (s, 1H, 10-H), 5.64 (d, J=7.0 Hz, 1H, 2-H), 4.96 (dd, J=9.5, 1.5 Hz, 1H, 5-H), 4.83 (br dd, J=12.5, 7.5 Hz, 1H, 13-H), 4.49 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.28 (d, J=8.0 Hz, 1H, 20-H), 4.15 (d, J=8.0 Hz, 1H, 20-H), 3.89 (d, J=7.0 Hz, 1H, 3-H), 2.53 (ddd, J=14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.30-2.20 (m, 2H, 14-CH2), 2.28 (s, 3H, Me), 2.19 (d, J=1.0 Hz, 3H, 18-Me), 2.18 (s, 3H, Me), 1.87 (ddd, J=14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.68 (s, 3H, Me), 1.63 (br s, 2H, OH, OH), 1.19 (s, 3H, Me), 1.04 (s, 3H, Me), 0.92 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.64-0.51 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 834.2270, M+Cs30 calcd for C36 H51 O11 NSi 834.2286.
DiTES taxoid 43. To a solution of alcohol 42 (12.6 mg, 0.018 mmol, previously azeotroped twice with benzene) and β-lactam 24 (17.0 mg, 0.0446 mmol, previously azeotroped twice with benzene) in THF (0.97 mL) at 0°C, prepared from the Ojima-Holton protocol (Holton, R. A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M.; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M.; Zucco, M.; Park, Y. H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), was added NaN(SiMe3)2 (0.054 mL of a 1.0 M solution in THF, 0.054 mmol) dropwise. The resulting solution was stirred for 0.5 h and poured into a mixture of ethylacetate (10 mL) and aqueous NH4 Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×5 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 50→95% ethylacetate in hexanes) to give 42 (1.0 mg, 8%) and 43 (8.6 mg, 48% based on 92% conversion) as a white solid.
Physical Data for DiTES taxoid 43. Rf =0.40 (silica , 50% ethylacetate in hexanes); IR (film) νmax 3433, 2955, 2880, 1730, 1662, 1370, 1238, 1112, 1018, 985, 824, 740 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 9.34 (d, J=2.0 Hz, 1H, pyridine), 8.82 (dd, J=5.0, 2.0 Hz, 1H, pyridine), 8.42 (ddd, J=8.0, 2.0, 2.0 Hz, 1H, pyridine), 7.74-7.69 (m, 2H), 7.51-7.20 (band, 9H), 7.08 (d, J=9.0 Hz, 1H, NH), 6.46 (s, 1H, 10-H), 6.22 (br t, J=9.0 Hz, 1H, 13-H), 5.74-5.66 (m, 2H, 2-H, 3'-H), 4.95 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.70 (d, J=2.0 Hz, 1H, 2'-H), 4.48 (dd, J=10.5,6.5 Hz, 1H, 7-H), 4.30 (d, J=8.0 Hz, 1H, 20-H), 4.21 (d, J=8.0 Hz, 1H, 20-H), 3.86 (d, J=7.0 Hz, 1H, 3-H), 2.58-2.48 (m, 1H, 6-H), 2.54 (s, 3H, Me), 2.40 (dd, J=15.5, 9.0 Hz, 1H, 14-H), 2.17 (s, 3H, Me), 2.14 (dd, J=15.5, 9.0 Hz, 1H, 14-H), 2.03 (br s, 3H, Me), 1.95-1.86 (m, 1H, 6-H), 1.73 (s, 4H, Me, OH), 1.22 (s, 3H, Me), 1.18 (s, 3H, Me), 0.93 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.82 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.65-0.37 (band, 12H, OSi(CH2 CH3)3, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 1215.4066, M+Cs30 calcd for C58 H78 O14 N2 Si2 1215.4046.
Taxoid 44. A solution of silyl ether 43 (6.4 mg, 0.0059 mmol) in THF (0.4 mL) at 25°C was treated with HF•pyridine (0.160 mL) and stirred for 1.25 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHCO3 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, ethylacetate) to give 44 (3.8 mg, 75%) as a colorless film.
Physical Data for Taxoid 44. Rf =0.59 (silica, ethylacetate); IR (film) νmax 3396, 2928, 1728, 1644, 1371, 1273, 1241, 1111, 1071 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 9.34 (br s, 1H, pyridine), 8.83 (br d, J=3.5 Hz, 1H, pyridine), 8.41 (br d, J=8.0 Hz, 1H, pyridine), 7.75-7.68 (m, 2H), 7.53-7.34 (band, 9H), 6.91 (d, J=9.0 Hz, 1H, NH), 6.27 (s, 1H, 10-H), 6.23 (br t, J=9.0 Hz, 1H, 13-H), 5.78 (dd, J=9.0, 2.5 Hz, 1H, 3'-H), 5.69 (d, J=7.0 Hz, 1H, 2-H), 4.95 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.79 (dd, J=5.5, 2.5 Hz, 1H, 2'-H), 4.41 (ddd, J=11.0, 6.5, 4.0 Hz, 1H, 7-H), 4.29 (d, J=8.5 Hz, 1H, 20-H), 4.20 (d, J=8.5 Hz, 1H, 20-H), 3.82 (d, J=7.0 Hz, 1H, 3-H), 3.54 (d, J=5.5 Hz, 1H, 2'-OH), 2.56 (ddd, J=14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.49 (d, J=4.0 Hz, 1H, 7-OH), 2.43-2.26 (m, 2H, 14-CH2), 2.38 (s, 3H, Me), 2.24 (s, 3H, Me), 1.89 (ddd, J=14.5, 11.0, 2.0 Hz, 1H, 6-H), 1.83 (s, 1H, OH), 1.82 (s, 3H, Me), 1.69 (s, 3H, Me), 1.25 (s, 3H, Me), 1.14 (s, 3H, Me); FAB HRMS (NBA/CsI) m/e 987.2325, M+Cs30 calcd for C46 H50 O14 N2 987.2316.
##STR30##
Acetate 45. A solution of alcohol 18 (50.0 mg, 0.0714 mmol) and 4-dimethylaminopyridine (DMAP, 26.0 mg, 0.213 mmol) in CH2 Cl2 (3.0 mL) at 25°C was treated with acetic anhydride (0.250 mL, 2.65 mmol) and stirred for 2.5 h. The reaction mixture was diluted with CH2 Cl2 (10 mL), treated with aqueous NaHCO3 (7 mL), and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 10% ethylacetate in benzene) to give 45 (41.0 mg, 77%) as an amorphous solid.
Physical Data for Acetate 45. Rf =0.27 (silica, 35% ethylacetate in hexanes); IR (film) νmax 3425, 2945, 1722, 1674, 1605, 1365, 1275, 1232, 1179, 1094; 1 H NMR (500 MHz, CDCl3) δ 7.89 (d, J=9.0 Hz, 2H, Ar), 6.64 (d, J=9.0 Hz, 2H, Ar), 6.56 (s, 1H, 10-H), 5.64 (d, J=6.5 Hz, 1H, 2-H), 4.90 (br d, J=8.0 Hz, 1H, 5-H), 4.45 (dd, J=10.5, 7.0 Hz, 1H, 7-H), 4.36 (d, J=9.0 Hz, 1H, 20-H), 4.11 (d, J=9.0 Hz, 1H, 20-H), 3.85 (d, J=6.5 Hz, 1H, 3-H), 3.05 (s, 6H, NMe2), 2.90 (d, J=20.0 Hz, 1H, 14-H), 2.62 (d, J=20.0 Hz, 1H, 14-H), 2.51 (ddd, J=14.0, 8.0, 7.0, 1H, 6-H), 2.20 (s, 3H, Me), 2.16 (s, 3H, Me), 2.15 (s, 3H, Me), 2.04 (s, 1H, OH), 1.84 (ddd, J=14.0, 10.5, 2.0 Hz, 1H, 6-H), 1.63 (s, 3H, Me), 1.23 (s, 3H, Me), 1.16 (s, 3H, Me), 0.89 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.58-0.53 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 874.8589, M+Cs30 calcd for C39 H55 O11 NSi 874.8594.
Alcohol 46. A solution of enone 45 (40.0 mg, 0.0539 mmol) in MeOH-THF (5.8:1, 4.1 mL) at 0°C was treated with NaBH4 (30.2 mg, 0.80 mmol, added by portions), stirred for 1 h, allowed to warm to 25° C. and stirred for 1.5 h. The reaction mixture was diluted with CH2 Cl2 (15 mL), treated with aqueous NH4 Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 25→50% ethylacetate in petroleum ether) to give 45 (6.0 mg, 15%) and 46 (30.0 mg, 88% based on 85% conversion) as an amorphous solid.
Physical Data for Alcohol 46. Rf =0.30 (silica, 50% ethylacetate in petroleum ether); 1 H NMR (500 MHz, CDCl3) δ 7.93 (d, J=9.0 Hz, 2H, Ar), 6.64 (d, J=9.0 Hz, 2H, Ar), 6.42 (s, 1H, 10-H), 5.57 (d, J=7.0 Hz, 1H, 2-H), 4.94 (br d, J=8.0 Hz, 1H, 5-H), 4.83-4.75 (m, 1H, 13-H), 4.46 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.34 (d, J=8.5 Hz, 1H, 20-H), 4.13 (d, J=8.5 Hz, 1H, 20-H), 3.82 (d, J=7.0 Hz, 1H, 3-H), 3.04 (s, 6H, Me2 N), 2.54-2.44 (m, 1H, 6-H), 2.26 (s, 3H, Me), 2.23 (d, J=7.5 Hz, 2H, 14-CH2), 2.16 (s, 6H, Me, Me), 2.08 (d, J=4.5 Hz, 1H, OH), 1.89-1.80 (m, 2H, 6-H, OH), 1.64 (s, 3H, Me), 1.16 (s, 3H, Me), 1.01 (s, 3H, Me), 0.89 (t, J=8.5 Hz, 9H, OSi(CH2 CH3)3), 0.62-0.48 (band, 6H, OSi(CH2 CH3)3).
DiTES taxoid 47. To a solution of alcohol 46 (14.0 mg, 0.0188 mmol, previously azeotroped twice with benzene) and β-lactam 24 (25.0 mg, 0.0656 mmol, previously azeotroped twice with benzene) in THF (0.75 mL) at 0°C, prepared from the Ojima-Holton protocol (Holton, R. A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M.; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M.; Zucco, M.; Park, Y. H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), was added NaN(SiMe3)2 (0.056 mL of a 1.0 M solution in THF, 0.056 mmol) dropwise. The resulting solution was stirred for 20 min and poured into a mixture of CH2 Cl2 (10 mL) and aqueous NH4 Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×5 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 10→15% ethylacetate in benzene, then 50% ethylacetate in petroleum ether) to give 47 (12.0 mg, 57%) as a white solid.
Physical Data for DiTES taxoid 47. Rf =0.26 (silica, 15% ethylacetate in PhH); IR (film) νmax 3425, 2946, 2882, 1722, 1669, 1600, 1365, 1275, 1238, 1179, 1094 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 7.96 (d, J=9.0 Hz, 2H, Ar), 7.77-7.72 (m, 2H), 7.54-7.26 (band, 8H), 7.12 (d, J=8.5 Hz, 1H, NH), 6.69 (d, J=9.0 Hz, 2H), 6.43 (s, 1H, 10-H), 6.23 (br t, J=9.0 Hz, 1H, 13-H), 5.68-5.63 (m, 2H, 2-H, 3'-H), 4.93 (br d, J=8.0 Hz, 1H, 5-H), 4.67 (d, J=2.0 Hz, 1H, 2'-H), 4.45 (dd, J=10.5 Hz, 6.5 Hz, 1H, 7-H), 4.36 (d, J=8.5 Hz, 1H, 20-H), 4.20 (d, J=8.5 Hz, 1H, 20-H), 3.78 (d, J=7.0 Hz, 1H, 3-H), 3.04 (s, 6H, Me2 N), 2.55-2.46 (m, 1H, 6-H), 2.53 (s, 3H, OAc), 2.36 (dd, J=15.5, 9.0 Hz, 1H, 14-H), 2.15 (s, 3H, Me), 2.09 (dd, J=15.5, 9.0 Hz, 1H, 14-H), 2.00 (d, J=1.0 Hz, 3H, Me), 1.92-1.84 (m, 2H, 6-H, OH), 1.67 (s, 3H, Me), 1.20 (s, 3H, Me), 1.16 (s, 3H, Me), 0.90 (t, J=8.0 Hz, 9H, OSi(CH2 CH33), 0.79 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.63-0.35 (band, 12 H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 1257.4503, M+Cs30 calcd for C61 H84 O14 N2 Si2 1257.4515.
Taxoid 48. A solution of silyl ether 47 (12.0 mg, 0.0107 mmol) in THF (1.0 mL) at 25°C was treated with HF•pyridine (0.05 mL) and stirred for 1.5 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHCO3 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 50→75% ethylacetate in petroleum ether) to give 48 (8.0 mg, 84%) as a colorless film.
Physical Data for Taxoid 48. Rf =0.44 (silica, 75% ethylacetate in petroleum ether); IR (film) νmax 3414, 2914, 2850, 1722, 1664, 1660, 1371, 1275, 1243, 1179 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 7.95 (d, J=9.0 Hz, 2H), 7.77-7.72 (m, 2H), 7.55-7.30 (band, 8H), 7.03 (d, J=9.0 Hz, 1H, NH), 6.67 (d, J=9.0 Hz, 2H), 6.24 (s, 1H, 10-H), 6.20 (br t, J=9.0 Hz, 1H, 13-H), 5.76 (dd, J=9.0, 2.5 Hz, 1H, 3'-H), 5.62 (d, J=7.0 Hz, 1H, 2-H), 4.93 (br d, J=7.5 Hz, 1H, 5-H), 4.76 (dd, J=5.0, 2.5 Hz, 1H, 2'-H), 4.37 (ddd, J=11.5, 6.5, 4.0 Hz, 1H, 7-H), 4.34 (d, J=8.5 Hz, 1H, 20-H), 4.18 (d, J=8.5 Hz, 1H, 20-H), 3.73 (d, J=7.0 Hz, 1H, 3-H), 3.57 (d, J=5.0 Hz, 1H, 2'-OH), 3.04 (s, 6H, Me2 N), 2.58-2.48 (m, 1H, 6-H), 2.44 (d, J=4.0 Hz, 1H, 7-OH), 2.37 (s, 3H, Me), 2.30-2.25 (m, 2H, 14-CH2), 2.22 (s, 3H, Me), 1.95 (s, 1H, OH), 1.88-1.81 (m, 1H, 6-H), 1.74 (d, J=1.0 Hz, 3H, Me), 1.65 (s, 3H, Me), 1.21 (s, 3H, Me), 1.11 (s, 3H, Me); FAB HRMS (NBA/CsI) m/e 1029.2760, M+Cs30 calcd for C49 H56 N2 O14 1029.2786.
##STR31##
Acetate 49. A solution of previous alcohol 19 and 4-dimethylaminopyridene (DMAP, 100 mg, 0.819 mmol in CH2 Cl2 (3 mL) at 25°C was treated with acetic anhydride (0.50 mL, 5.30 mmol and stirred for 3 h. The reaction mixture was diluted with CH2 Cl2 (5 mL), treated with aqueous NaHCO3 (7 mL, and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×10 mL. The combined organic layer was washed with brine (5 mL, dried (MgSO4), concentrated, and purified by preparative TLC (silica, 10% ethylacetate in benzene to give 49 (54.1 mg, 89% from carbonate 7) as an amorphous solid.
Physical data for Acetate 49. Rf =0.27 (20% ethylacetate in petroleum ether); IR (film) vmax 3416, 2953, 2879, 1726, 1676, 1370, 1224, 1089 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.66 (s, 1H, naphthalene), 8.06 (dd, 1H, J=9.0, 2.0 Hz naphthalene), 7.98-7.89 (m, 3H, naphthalene), 7.68-7.55 (m, 2H, naphthalene), 6.61 (s, 1H, 10-H), 5.75 (d, J=7.0 Hz, 1H, 2-H), 4.95 (br d, J=8.0 Hz, 1H, 5-H), 4.50 (dd, J=10.5, 7.0 Hz, 1H, 7-H), 4.35 (d, J=8.5 Hz, 1H, 20-H), 4.16 (d, J=8.5 Hz, 1H, 20-H), 3.96 (d, J=8.5 Hz, 1H, 20-H), 3.96 (d, J=7.0 Hz, 1H, 3-H), 3.03 (d, J=20.0 Hz, 1H, 14-H), 2.70 (d, J=20.0 Hz, 1H, 14-H), 2.61-2.50 (m, 2H, 6-H, OH), 2.27 (s, 3H, Me), 2.24 (s, 3H, Me), 2.21 (s, 3H, Me), 1.91-1.83 (m, 1H, 6-H), 1.70 (s, 3H, Me), 1.30 (s, 3H, Me), 1.20 (s, 3H, Me), 0.93 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.66-0.57 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 881.2326, M+Cs30 calcd for C41 H52 O11 Si 881.2333.
Alcohol 50. A solution of enone 49 (54.1 mg, 0.0722 mmol) in MeOH (10 mL) at 25°C was treated with NaBH4 (54.5 mg, 1.44 mmol, added by portions) and stirred for 2.0 h. The reaction mixture was diluted with CH2 Cl2 (10 mL), treated with aqueous NH4 Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 20% ethylacetate in petroleum ether) to give 50 (26 mg, 48%) as an amorphous solid.
Physical Data for Alcohol 50. Rf =0.12 (20% ethylacetate in petroleum ether); IR (film) νmax 3524, 2953, 1719, 1369, 1231, 1093, 829 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.70 (s, 1H, naphthalene), 8.11 (dd, J=8.5, 1.5 Hz, 1H, naphthalene), 7.96-7.86 (m, 3H, naphthalene), 7.65-7.54 (m, 2H, naphthalene), 6.45 (s, 1H, 10-H), 5.68 (d, J=7.0 Hz, 1H, 2-H), 4.98 (br d, J=8.0 Hz, 1H, 5-H), 4.88-4.81 (m, 1H, 13-H), 4.51 (dd, J=10.5, 7.0 Hz, 1H, 7-H), 4.34 (d, J=8.5 Hz, 1H, 20-H), 4.19 (d, J=8.5 Hz, 1H, 20-H), 3.93 (d, J=7.0 Hz, 1H, 3-H), 2.58-2.50 (m, 1H, 6-H), 2.41-2.14 (m, 3H, 14-CH2, 13-OH), 2.37 (s, 3H, Me), 2.21 (br s, 3H, Me), 2.19 (s, 3H, Me), 1.92-1.84 (m, 1H, 6-H), 1.72 (s, 1H, OH) 1.71 (s, 3H, Me), 1.22 (s, 3H, Me), 1.05 (s, 3H, Me), 0.93 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.65-0.51 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 883.2484, M+Cs30 calcd for C41 H54 O11 Si 883.2490.
DiTES taxoid 51. To a solution of alcohol 50 (20.0 mg, 0.0266 mmol, previously azeotroped twice with benzene) and β-lactam 24 (20.0 mg, 0.0525 mmol, previously azeotroped twice with benzene) in THF (1.1 mL) at -78°C, prepared from the Ojima-Holton protocol (Holton, R. A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M.; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M.; Zucco, M.; Park, Y. H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), was added NaN(SiMe3)2 (0.065 mL of a 1.0 M solution in THF, 0.065 mmol) dropwise. The resulting solution was stirred for 10 min and poured into a mixture of CH2 Cl2 (10 mL) and aqueous NH4 Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×5 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 20% ethylacetate in petroleum ether) to give 51 (18.7 mg, 62%) as a white solid.
Taxoid 52. A solution of silyl ether 51 (18.7 mg, 0.0165 mmol) in THF (2 mL) at 25°C was treated with HF•pyridine (1 mL) and stirred for 1 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHCO3 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 50% ethylacetate in petroleum ether) to give 52 (12.8 mg, 86%) as a colorless film.
Physical Data for Taxoid 52. Rf =0.16 (silica, 50% ethylacetate in petroleum ether); IR (film) νmax 3420, 2967, 2896, 1721, 1652, 1519, 1370, 1233, 1073, 776 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 8.67 (s, 1H, naphthalene), 8.04 (dd, J=8.5, 1.5 Hz, 1H, naphthalene), 7.95 (br d, J=8.5 Hz, 1H, naphthalene), 7.87 (br d, J=9.0 Hz, 1H), 7.81 (br d, J=8.5 Hz, 1H), 7.65-7.61 (m, 2H), 7.56-7.51 (m, 1H), 7.49-7.22 (band, 9H), 6.94 (d, J=9.0 Hz, 1H, NH), 6.23-6.16 (m, 2H, 10-H, 13-H), 5.78 (dd, J=9.0, 2.0 Hz, 1H, 3'-H), 5.64 (br d, J=7.0 Hz, 1H, 2-H), 4.87 (br d, J=8.0 Hz, 1H, 5-H), 4.78-4.72 (m, 1H, 2'-H), 4.38-4.31 (m, 1H, 7-H), 4.24 (d, J=8.5 Hz, 1H, 20-H), 4.16 (d, J=8.5 Hz, 1H, 20-H), 3.76 (d, J=7.0 Hz, 1H, 3-H), 3.53 (br s, 1H, OH), 2.52-2.43 (m, 1H, 6-H), 2.42 (d, J=4.0 Hz, 1H, OH), 2.40 (s, 3H, Me), 2.36 (dd, J=15.5, 9.0 Hz, 1H, 14-H), 2.25 (dd, J=15.5, 9.0 Hz, 1H, 14-H), 2.17 (s, 3H, Me), 1.85-1.77 (m, 2H, 6-H, OH), 1.74 (br s, 3H, Me), 1.63 (s, 3H, Me), 1.17 (s, 3H, Me), 1.09 (s, 3H, Me); FAB HRMS (NBA/CsI) m/e 1036.2505, M+Cs30 calcd for C51 H53 NO14 1036.2520
##STR32##
Acetate 53. A solution of alcohol 23 (25.2 mg, 0.0351 mmol) and 4-dimethylaminopyridine (DMAP, 12.2 mg, 0.0999 mmol) in CH2 Cl2 (1.5 mL) at 25°C was treated with acetic anhydride (0.120 mL, 1.27 mmol) and stirred for 1.5 h. The reaction mixture was diluted with CH2 Cl2 (5 mL), treated with aqueous NaHCO3 (7 mL), and stirred vigorously for 25 min. The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 30% ethylacetate in petroleum ether) to give 53 (25.3 mg, 95%) as a colorless oil.
Physical Data for Acetate 53. Rf =0.41 (silica, 10% ethylacetate in benzene, 2 elutions); IR (film) νmax 3471, 2954, 2881, 1729, 1675, 1370, 1226, 986, 824, 738 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 7.38-7.25 (band, 5H, SPh), 6.54 (s, 1H, 10-H), 5.49 (br d, J=6.5 Hz, 1H, 2-H), 4.90 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.42 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.37 (d, J=8.0 Hz, 1H, 20-H), 4.17 (d, J=8.0 Hz, 1H, 20-H), 3.78 (d, J=6.5 Hz, 1H, 3-H), 3.23-3.13 (m, 2H, CH2 SPh), 2.78 (d, J=20.0 Hz, 1H, 14-H), 2.72-2.58 (m, 3H, CH2 CH2 SPh, 14-H), 2.52 (ddd, J=14.5, 9.5, 6.5, 1H, 6-H), 2.45 (s, 1H, OH), 2.21 (s, 3H, Me), 2.15 (s, 3H, Me), 2.04 (s, 3H, Me), 1.86 (ddd, J=14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.62 (s, 3H, Me), 1.23 (s, 3H, Me), 1.19 (s, 3H, Me), 0.91 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.64-0.52 (band, 6H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 891.2225, M+Cs30 calcd for C39 H54 O11 SSi 891.2210.
Alcohol 54. A solution of enone 53 (24.4 mg, 0.032 mmol) in MeOH-THF (5:1, 1.9 mL) at 0°C was treated with NaBH4 (18.1 mg, 0.48 mmol, added by portions) and stirred for 1.25 h. The reaction mixture was diluted with CH2 Cl2 (5 mL), treated with aqueous NH4 Cl (5 mL), and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×5 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 30% ethylacetate in hexanes) to give 54 (14.6 mg, 60%) as an amorphous solid.
Physical Data for Alcohol 54. Rf =0.11 (silica, 30% ethylacetate in hexanes); IR (film) νmax 3487, 2938, 2880, 1729, 1586, 1369, 1234, 977, 738 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 7.40-7.23 (band, 5H, SPh), 6.42 (s, 1H, 10-H), 5.43 (d, J=7.0 Hz, 1H, 2-H), 4.94 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.85-4.78 (m, 1H, 13-H), 4.43 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.37 (d, J=8.0 Hz, 1H, 20-H), 4.18 (d, J=8.0 Hz, 1H, 20-H), 3.74 (d, J=7.0 Hz, 1H, 3-H), 3.25-3.15 (m, 2H, CH2 SPh), 2.71-2.57 (m, 2H, CH2 CH2 SPh), 2.51 (ddd, J=14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.25 (dd, J=15.5, 9.5 Hz, 1H, 14-H), 2.16 (s, 3H, Me), 2.15 (d, J=1.0 Hz, 3H, 18-Me), 2.15 (s, 3H, Me), 2.09 (dd, J=15.5, 7.0 Hz, 1H, 14-H), 2.05 (br s, 1H, OH), 1.99-1.96 (m, 1H, OH), 1.86 (ddd, J=14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.63 (s, 3H, Me), 1.15 (s, 3H, Me), 1.04 (s, 3H, Me), 0.91 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.64-0.50 (band, 6H, Si(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e 893.2350, M+Cs30 calcd for C39 H56 O11 SSi 893.2367.
DiTES taxoid 55. To a solution of alcohol 54 (21.8 mg, 0.0286 mmol, previously azeotroped twice with benzene) and β-lactam 24 (33.0 mg, 0.0866 mmol, previously azeotroped twice with benzene) in THF (1.1 mL) at 0°C, prepared from the Ojima-Holton protocol (Holton, R. A. Chem Abstr. 1990, 114, 164568q; Ojima, I.; Habus, I.; Zhao, M.; Georg, G. I.; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-1683; Ojima, I.; Habus, I.; Zhao, M.; Zucco, M.; Park, Y. H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), was added NaN(SiMe3)2 (0.086 mL of a 1.0 M solution in THF, 0.086 mmol) dropwise. The resulting solution was stirred for 20 min and poured into a mixture of CH2 Cl2 (10 mL) and aqueous NH4 Cl (5 mL). The organic layer was separated and the aqueous layer was extracted with CH2 Cl2 (2×5 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by flash chromatography (silica, 15→30→50% ethylacetate in petroleum ether) to give 55 (13.8 mg, 42%) as an amorphous solid.
Physical Data for DiTES taxoid 55. Rf =0.40 (silica, 30% ethylacetate in hexanes); IR (film) νmax 3437, 2952, 2879, 1735, 1662, 1482, 1369, 1236, 1128, 981, 740 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 7.82-7.76 (m, 2H), 7.54-7.16 (band, 13H), 7.11 (d, J=9.0 Hz, 1H, NH), 6.41 (s, 1H, 10-H), 6.18 (br t, J=9.0 Hz, 1H, 13-H), 5.62 (dd, J=9.0, 2.0 Hz, 1H, 3'-H), 5.49 (d, J=7.0 Hz, 1H, 2-H), 4.93 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.64 (d, J=2.0 Hz, 1H, 2'-H), 4.42 (dd, J=10.5, 6.5 Hz, 1H, 7-H), 4.40 (d, J=8.0 Hz, 1H, 20-H), 4.21 (d, J=8.0 Hz, 1H, 20-H), 3.70 (d, J=7.0 Hz, 1H, 3-H), 3.23-3.17 (m, 2H, CH2 SPh), 2.78-2.69 (m, 1H, HCHCH2 SPh), 2.67-2.57 (m, 1H, HCHCH2 SPh), 2.55-2.46 (m, 2H, 6-H, OH), 2.38 (s, 3H, Me), 2.27-2.10 (m, 2H, 14-CH2), 2.16 (s, 3H, Me), 1.98 (d, J=1.0 Hz, 3H, Me), 1.89 (ddd, J=14.0, 11.0, 2.0 Hz,
1H, 6-H), 1.64 (s, 3H, Me), 1.18 (s, 3H, Me), 1.17 (s, 3H., Me), 0.91 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.81 (t, J=8.0 Hz, 9H, OSi(CH2 CH3)3), 0.64-0.36 (band, 12 H, OSi(CH2 CH3)3); FAB HRMS (NBA/CsI) m/e M+Cs+ 1274.4125 calcd for C61 H83 O14 SSi2 1274.4127.
Taxoid 56. A solution of silyl ether 55 (8.1 mg, 0.0071 mmol) in THF (0.5 mL) at 25°C was treated with HF•pyridine (0.150 mL) and stirred for 3.75 h. The reaction mixture was poured into a mixture of ethylacetate (10 mL) and aqueous NaHCO3 (5 mL) and the resulting mixture was stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with ethylacetate (2×10 mL). The combined organic layer was washed with brine (5 mL), dried (MgSO4), concentrated, and purified by preparative TLC (silica, 60% ethylacetate in petroleum ether) to give 56 (3.2 mg, 49%) as a colorless film.
Physical Data for Taxoid 56. Rf =0.39 (silica, 60% ethylacetate in petroleum ether); IR (film) νmax 3426, 2928, 1731, 1642, 1371, 1238, 1070, 739, 709 cm-1 ; 1 H NMR (500 MHz, CDCl3) δ 7.80-7.75 (m, 2H), 7.55-7.18 (band, 13 H), 6.94 (d, J=9.0 Hz, 1H, NH), 6.23 (s, 1H, 10-H), 6.19 (br t, J=9.0 Hz, 1H, 13-H), 5.74 (dd, J=9.0, 2.5 Hz, 1H, 3'-H), 5.47 (d, J=7.0 Hz, 1H, 2-H), 4.93 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.74 (dd, J=5.0, 2.5 Hz, 1H, 2'-H), 4.38 (d, J=8.0 Hz, 1H, 20-H), 4.35 (ddd, J=11.0, 6.5 Hz, 4.5 Hz, 1H, 7-H), 4.21 (d, J=8.0 Hz, 1H, 20-H), 3.67 (d, J=7.0 Hz, 1H, 3-H), 3.51 (d, J=5.0 Hz, 1H, 2'-OH), 3.28-3.14 (m, 2H, CH2 SPh), 2.77-2.68 (m, 1H, HCHCH2 SPh), 2.67-2.59 (m, 1H, HCHCH2 SPh), 2.54 (ddd, J=14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.44 (d, J=4.5 Hz, 1H, 7-OH), 2.36 (dd, J=15.5, 9.0 Hz, 1H, 14-H), 2.26 (br s, 1H, OH), 2.23 (s, 3H, Me), 2.21 (s, 3H, Me), 2.18 (dd, J=15.5, 9.0 Hz, 1H, 14-H), 1.88 (ddd, J=14.5, 11.0, 2.0 Hz, 1H, 6-H), 1.75 (d, J=1.0 Hz, 3H, Me), 1.63 (s, 3H, Me), 1.24 (s, 3H, Me), 1.10 (s, 3H, Me); FAB HRMS (NBA/CsI) m/e 1046.2410, M+Cs30 calcd for C49 H55 O14 NS 1046.2398.
##STR33##
MPA taxoid 57. A solution of taxoid 36 (4.3 mg, 0.005 mmol) and triethylamine (0.0033 mL, 0.0237 mmol) in CH2 Cl2 (0.2 mL) at 25°C was treated with 2-fluoro-1-methylpyridinium p-toluenesulfonate (2.1 mg, 0.0075 mmol) and stirred for 35 min. The clear colorless solution rapidly turned to a clear pale yellow. The course of the reaction was monitored through thin layer chromatography (TLC)(E. Merck RP-18 silica, 65 tetrahydrofuran: 35 water, UV/phospho-molybidic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC. The reaction mixture was directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give 36 (0.8 mg, 19%) and 57 (4.1 mg, 100% based on 81% conversion) as a colorless film.
Physical Data for taxoid 57 1 H NMR (500 MHz, CDCl3) δ 10.5 (d, J=7.5 Hz, 1H), 8.44 (ddd, J=9.0, 7.5, 2.0 Hz, 1H), 8.33-8.29 (m, 2H), 8.15 (dd, J=3.0, 1.0 Hz, 1H, thiophene), 8.12 (br d, J=6.0 Hz, 1H), 7.84 (br d, J=8.5 Hz, 1H), 7.74-7.69 (m, 2H), 7.53 (dd, J=5.0, 1.0 Hz, 1H, thiophene), 7.48-7.34 (band, 7H), 7.16-7.12 (m, 1H), 6.53-6.43 (m, 1H, 2'-H), 6.21 (s, 1H, 10-H), 6.03 (dd, J=10.5, 8.0 Hz, 1H, 3'-H), 5.82 (br t, J=9.0 Hz, 1H, 13-H), 5.44 (d, J=7.0 Hz, 1H, 2-H), 4.90 (dd, J=9.5, 2.0 Hz, 1H, 5-H), 4.33 (dd, J=11.0, 6.5 Hz, 1H, 7-H), 4.30 (d, J=8.0 Hz, 1H, 20-H), 4.15 (d, J=8.0 Hz, 1H, 20-H), 4.08 (s, 3H, N+ Me), 3.68 (d, J=7.0 Hz, 1H, 3-H), 2.58-2.49 (m, 1H, 6-H), 2.52 (s, 3H, OAc), 2.21 (s, 3H, OAc), 2.04 (s, 3H, OAc), 2.02 (br s, 2H, OH, OH), 1.88 (ddd, J=14.5, 11.5, 2.0 Hz, 1H, 6-H), 1.78 (br s, 3H, 18-Me), 1.64 (s, 3lH, Me), 1.61 (dd, J=16.0, 7.0 Hz, 1H, 14-H), 1.18 (dd, J=16.0, 9.0 Hz, 1H, 14-H), 1.13 (s, 3 H, Me), 1.08 (s, 3 H, Me).
##STR34##
MPA taxoid 58. A solution of taxoid 32 (1.0 equiv.) and triethylamine (4.7 equiv.) in CH2 Cl2 (0.025 M) at 25°C is treated with 2-fluoro-1-methylpyridinium p-toluenesulfonate from Aldrich Chemical company inc. (1.5 equiv.) and stirred for 35 minutes. The course of the reaction was monitored through thin layer chromatography (TLC)(E. Merck RP-18 silica, 65 tetrahydrofuran: 35 water, UV/phospho-molybidic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give 58 as a colorless film.
PACPreparation of MPA Taxoid 59 ##STR35##
MPA taxoid 59. The synthesis of the taxoid-7-MPA 59 differs only slightly from the synthesis of taxoid-2'-MPA 58. The C-2 taxoid 32 is dissolved in methylene chloride (0.006 M) and treated sequentially with triethylamine (40 equivalents) and 2-fluoro-1-methyl-pyridinium tosylate (10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 5 minutes. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give 59 as a colorless film.
##STR36##
MPA taxoid 60. The synthesis of the taxoid-7-MPA 60 differs only slightly from the synthesis of taxoid-2'-MPA 57. The C-2 taxoid 36 is dissolved in methylene chloride (0.006 M) and treated sequentially with triethylamine (40 equivalents) and 2-fluoro-1-methyl-pyridinium tosylate (10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 5 minutes. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give 60 as a colorless film.
##STR37##
C-2-taxoid-2'-onium salts 62-66. A solution of taxoid (62-66) I. (1.0 equiv.) and triethylamine (4.7 equiv.) in CH2 Cl2 (0.025 M) at 25°C is treated with 2-fluoro-1-methylpyridinium p-toluenesulfonate from Aldrich Chemical company inc. (1.5 equiv.) and stirred for 35 minutes. The course of the reaction was monitored through thin layer chromatography (TLC)(E. Merck RP-18 silica, 65 tetrahydrofuran: 35 water, UV/phospho-molybidic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give (62-66) II. as a colorless film.
##STR38##
C-2-taxoid-7-onium salts 67-72. The synthesis of the taxoid-7-methyl-pyridinium salts (67-72) II, differs only slightly from the synthesis of taxoid-2'-methyl-pyridinium salts (62-66) II. The C-2 taxoid (67-72) I is dissolved in methylene chloride (0.006 M) and treated sequentially with triethylamine (40 equivalents) and 2-fluoro-1-methyl-pyridinium tosylate (10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 5 minutes. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give (67-72) II as a colorless film.
##STR39##
C-2-taxoid-bis-2',7-onium salts 73-80. The synthesis of C-2-taxoid-bis-2',7-methyl-pyridinium salts II (73-80), differs from the synthesis of taxoid-7-methyl-pyridinium salts (67-72) II only with respect to reaction time. The C-2 taxoid (73-80) I is dissolved in methylene chloride (0.006 M) and treated sequentially with triethylamine (40 equivalents) and 2-fluoro-1-methyl-pyridinium tosylate (10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 18 hours. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give (73-80) II as a colorless film.
##STR40##
C-2-taxoid-2'-benzothiazolium salts 81-88. A solution of taxoid 81-88 (1.0 equiv.) and triethylamine (4.7 equiv.) in CH2 Cl2 (0.025 M) at 25°C is treated with 2-fluoro-1-methylpyridinium p-toluenesulfonate from Aldrich Chemical company inc. (1.5 equiv.) and stirred for 35 minutes. The course of the reaction was monitored through thin layer chromatography (TLC)(E. Merck RP-18 silica, 65 tetrahydrofuran: 35 water, UV/phospho-molybidic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give 81-88 as a colorless film.
##STR41##
C-2-taxoid-7-benzothiazolium salts (89-96). The synthesis of the taxoid-7-benzothiazolium salts (89-96) II, differs only slightly from the synthesis of taxoid-2'-benzothiazolium salts (81-88) II. The C-2 taxoid (89-96) I is dissolved in methylene chloride (0.006 M) and treated sequentially with triethylamine (40 equivalents) and 2-fluoro-3-ethylbenzothiazolium tetrafluoroborate (10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 5 minutes. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give (89-96) II as a colorless film.
##STR42##
C-2-taxoid-2'-benzoxazolium salts 97-104. A solution of taxoid (97-104) I. (1.0 equiv.) and triethylamine (4.7 equiv.) in CH2 Cl2 (0.025 M) at 25°C is treated with 2-chloro-3-ethylbenzoxazolium tetrafluoroborate from Aldrich Company (1.5 equiv.) and stirred for 35 minutes. The course of the reaction was monitored through thin layer chromatography (TLC)(E. Merck RP-18 silica, 65 tetrahydrofuran: 35 water, UV/phospho-molybidic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give (97-104) II. as a colorless film.
##STR43##
C-2-taxoid-7-benzoxazolium salts (105-112). The synthesis of the taxoid-7-benzoxazolium salts (105-112) II, differs only slightly from the synthesis of taxoid-2'-benzoxazolium salts (97-104) II. The C-2 taxoid (105-112) I is dissolved in methylene chloride (0.006 M) and treated sequentially with triethylamine (40 equivalents) and 2-chloro-3-ethylbenzoxazolium tetrafluoroborate from Aldrich Company (10 equivalents) Aldrich Chemicals, and allowed to stir at ambient temperature for 5 minutes. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give (105-112) II as a colorless film.
##STR44##
C-2-taxoid-2'-pyrimidinium salts 113-120. A solution of taxoid (113-120) I. (1.0 equiv.) and triethylamine (4.7 equiv.) in CH2 Cl2 (0.025 M) at 25°C is treated with 2-chloro-methyl-pyrimidinium fluoride from Aldrich Company (1.5 equiv.) and stirred for 35 minutes. The course of the reaction was monitored through thin layer chromatography (TLC)(E. Merck RP-18 silica, 65 tetrahydrofuran: 35 water, UV/phospho-molybidic acid) and after thirty minutes of stirring at ambient temperature, judged complete as no taxol remained and only one compound was apparent by TLC. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give (113-120) II. as a colorless film.
##STR45##
C-2-taxoid-7-pyrimidinium salts (121-128). The synthesis of the taxoid-7-pyrimidinium salts (121-128) II, differs only slightly from the synthesis of taxoid-2'-pyrimidinium salts (113-120) II. The C-2 taxoid (121-128) I is dissolved in methylene chloride (0.006 M) and treated sequentially with triethylamine (40 equivalents) and 2-chloro-methyl-pyrimidinium fluoride from Aldrich Company (10 equivalents), and allowed to stir at ambient temperature for 5 minutes. The reaction mixture is then directly purified by HPLC (Vydak RP-18, 22.5×3 mm, A→B 0.5 h linear, A: 20% MeOH in 20 mM NH4 OAc, B: 100% MeOH, 9 mL/min, RT=26.12) to give (121-128) II as a colorless film.
Wrasidlo, Wolfgang, Nicolaou, K. C., Guy, Rodney K., Pitsinos, Emmanuel
| Patent | Priority | Assignee | Title |
| 6362217, | Mar 17 2000 | Bristol-Myers Squibb Company | Taxane anticancer agents |
| 8658600, | Jun 26 2009 | Brandeis University | Antineoplastic hydrogels, and enzyme-instructed preparations thereof |
| 9408921, | Jun 26 2009 | Brandels University | Antineoplastic hydrogels, and enzyme-instructed preparations thereof |
| Patent | Priority | Assignee | Title |
| 4942184, | Mar 07 1988 | UNITED STATES OF AMERICA, THE, AS REPRESENTED BY THE SECRETARY OF THE DEPARTMENT OF HEALTH AND HUMAN SERVICES | Water soluble, antineoplastic derivatives of taxol |
| 4960790, | Mar 09 1989 | University of Kansas | Derivatives of taxol, pharmaceutical compositions thereof and methods for the preparation thereof |
| 5274137, | Jun 23 1992 | SCRIPPS RESEARCH INSTITUTE, THE | Intermediates for preparation of taxols |
| 5422364, | Sep 04 1992 | SCRIPPS RESEARCH INSTITUTE, THE | Water soluble taxol derivatives |
| 5440057, | Aug 20 1993 | SCRIPPS RESEARCH INSTITUTE, THE | Access to taxol analogs |
| 5461169, | Jun 23 1992 | SCRIPPS RESEARCH INSTITUTE, THE | Total synthesis of taxol and taxol analogs |
| 5481007, | Jun 23 1992 | SCRIPPS RESEARCH INSTITUTE, THE | Taxoid synthesis |
| 5504222, | Aug 20 1993 | SCRIPPS RESEARCH INSTITUTE, THE | Transformations of taxol |
| 5608087, | Sep 04 1992 | The Scripps Research Institute | Water soluble taxol derivatives |
| 5731334, | Jan 11 1994 | SCRIPPS RESEARCH INSTITUTE, THE | Method for treating cancer using taxoid onium salt prodrugs |
| 5750691, | Aug 20 1993 | The Scripps Research Institute | Access to taxol analogs |
| 5760240, | Jun 23 1992 | The Scripps Research Institute | Taxoid synthesis |
| 5786489, | Feb 08 1994 | The Scripps Research Institute | Transformations of taxol |
| 5817840, | Sep 04 1992 | The Scripps Research Institute | Water soluble taxol derivatives |
| 6043382, | Aug 20 1993 | The Scripps Research Institute | Transformations of taxol |
| EP473326A1, | |||
| WO8908453, | |||
| WO9405282, |
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